The Fourth CJUA annual meeting
2010

China­Japan urological association

PROGRAM ＆ ABSTRACTS

DateOct. 16, 2010
Venue
XIAN GREENLAND PICO
International Convention & Exhibition Center Room 105
http://www.gpcec.com
CJUA Home­page

URL: http://www.uro.jp/nichu/index.html

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PROGRAM of the Fourth CJUA annual meeting(2010.10.16 Xian

8:30 to 8:40

China)

Opening Address

Prof. Yanqun Na, President of CJUA (China)
8:40 to 9:10

Special Lecture

Chairman: Prof. Hiromi Kumon, Okayama University
Comprehensive SNP Analyses on Genetic Predisposition for Incidence of 14 Major
Cancers in Japan: Special Reference to Urologic Cancers.
Prof. Kenji Shimizu, Department of Molecular Genetics, Okayama University
9:10 to 10:10

Session I (7 minutes presentation following 3 minutes discussion)

Chairmen: Prof. Yukio Homma, Tokyo, Japan
Prof. Xianghua Zhang, Beijing, China
1. Cooperative Research on Genetic Predisposition for prostate Cancer in Asia byAnalyzing Cumulative Association of SNPs ­The analyses data in Japan­.
Huang Peng, et al.

Okayama University, Japan.

2. Cooperative Research on Genetic Predisposition for prostate Cancer in Asia byAnalyzing Cumulative Association of SNPs ­The analyses data in Chinese­.
Lei Wang, et al.

Peking University People’s hospital, China

3. Complications of Urological Laparoscopic Surgery: A Single Institute Experienceof 1,017 Procedures.
Takaaki Inoue, et al. Kansai Medical University, Japan
4. Percutaneous renal surgery performance on live pigs can be improved bypracticing on a biologic porcine kidney model.
Yi Zhang, GangWang, Yanqun Na. Peking University Wu Jie Ping Urology Center,
China

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5. Primary congenital bladder diverticula in adult.
Wang Jianbo, Sun Xishuang. Dalian Medical University, China
6. High­intensity focused ultrasound as salvage therapy for patients with recurrent
prostate cancer after external beam radiation, brachytherapy or proton therapy.
Toyoaki Uchida et al.

Tokai University, Japan

10:10 ­ 10:30

Coffee Break

10:30 to 11:20

Session II(7 minutes presentation following 3 minutes discussion)

Chairmen: Prof. Toyoaki Uchida, Tokyo Japan
Prof. Yi Zhang, Beijing, China
7. Prognostic factors for renal cell carcinoma with bone metastasis: who are the longsurvivors?
Haruki Kume, et al. Tokyo University, Japan
8. The expression and clinical value of cytokeratin in peripheral blood of patientswith urinary carcinoma.
Xiaoqiang Liu, Sun Guang. Tianjin Medical University, China
9. Systemic T­cell activation following in situ gene therapy in prostate cancer
patients.
Shinji Kurosaka, Takefumi Satoh, Makoto Kubo, et al. Kitasato University, Japan
10. The expression and role of transient receptor potential channel A1 in the
bladders of rat and human being.
Shuqi Du, the First Affiliated Hospital, China Medical University, China
11. Synergistic apoptosis of lexatumumab and cisplatin against human renal cell
carcinoma cells in a caspase­dependent manner.
Xiuxian Wu, et al.
11:20 to 11:30

Kagawa University, Japan

Closing Address

Prof. Hiromi Kumon, President of CJUA (Japan)
11:30 ­ 12:30

CJUA Leadership Meeting3

Special Lecture
Comprehensive SNP Analyses on Genetic Predisposition for Incidence of 14 Major
Cancers in Japan: Special Reference to Urologic Cancers.
Kenji Shimizu
Department of Molecular Genetics, Okayama University Graduate School of
Medicine, Dentistry, and Pharmaceutical Sciences, Okayama, Japan
Our aim is to establish a comprehensive strategy for predicting individual’s genetic
predisposition for most major cancers in Japan through analyzing the overlap of
SNP­genotypes at risk on each individual for each cancer.
We have analyzed so far about 270 missense (ms) ­SNP with a minor allele
frequency (MAF) > 5%, from 225 cancer­related genes for differential distribution of
the genotypes between area­ and age­matched non­cancer controls (about 970
samples) and 14 kinds of cancer cases (in total, about 2,500 cases).
We have found 44 ms­SNPs from 40 genes are significantly associated withincidence of at least one of 14 kinds of major cancers, which altogether cover
approximately 80% of total cancer patients in Japan. Most notably, significant
implications of 31 out of these 44 SNP with any cancers were novel findings. In
respect to Urologic Cancers, in total of 14, 9, and 6 genes were found to be
associated with incidence of Prostate, Renal Cell, and Bladder Cancer, respectively.
Of particular interest, stratification analyses revealed that the repertoire ofsignificantly associating SNP genotypes were entirely different between genders(Lung, Colorectal, Pancreatic, and Kidney Cancers) and between age groups (by 70
years; Prostate Cancer, by 55 years; Breast Cancer).
As a way to assess the cumulative effects of the overlapping SNP genotypes at risk,
we introduced a measure, MOR (Multiplied Odds Ratio), which is the “product” ofeach OR value of the overlapping SNP genotypes on each person for each cancer
type. Possible bias by gene­gene interactions should be equally applied for controls
and cancer cases in case­control studies. Thus, the relative comparison of the MORvalue for a single kind of cancer should be substantially reliable for assessing the
cumulative risk in a relative sense, albeit the absolute MOR value may be lessmeaningful.
We have analyzed most of the specimens for most of the 44 SNP genotypes andMOR values were determined on each sample for each cancer. As expected, theMOR value was much higher in cancer cases than in control samples, mean valuesof which sometimes exceeded ~10 fold difference.

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According to the stratification of the MOR distribution for each cancer type andrisk­assessment by the likelihood ratio analysis and calculation of the lifetime
probability of each cancer incidence, we have found that about 50~80% of Japanese
population are relatively cancer­protective, 10~30% are intermediate, 5~20% arehigh­risk, and 0.5~2% are extremely high­risk on most of the major types of cancer.
For example, the highest risk group for Prostate Cancer (PSC) of > 70 years onset,
based on the overlap of 6 SNP genotypes, was classified as the population with MOR> 6, and constitute only 0.5% of healthy population and 15% of PSC cases, whereas
the lowest risk group with MOR < 1.5 occupy 57% of healthy population and 31% of
PSC cases. Taking the lowest risk group as the reference, the highest risk groupexerts about 64­fold increased risk for PSC ( OR = 64.2, 95% CI = 11.5­359, p­value:
2.28 x 10 ­8 ). Japanese males of this category are predicted to have 33% lifetime
probability for incidence of PSC (one in three).
Thus, our way of analysis of the genetic predisposition serves an efficient strategy
to predict the susceptibility to specific cancers on each person. We can make usethem to prevent cancer by altering life­styles and to detect early cancers by periodic
medical checking of the specified organs of the confined individuals at risk. This
strategy may eventually be applicable in East­Asian countries sharing similar
genetic background with Japanese and these trials are now underway.

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Abstract
1. Cooperative Research on Genetic Predisposition for prostate Cancer in Asia
by Analyzing Cumulative Association of SNPs ­The analyses data in Japan­
Peng Huang 1) 3), Haruki Kaku 1) 3), Akiko Sakai２),Masami Watanebe1) 3), Takashi
Saika1),Yasutomo Nasu1), Hiromi Kumon 1) 3) & Kenji Shimizu２)
1) Department of Urology,2) Department of Molecular Genetics,
3) The Center of Gene and Cell Therapy, Okayama University Graduate School of
Medicine, Dentistry and Pharmaceutical Sciences, Okayama 700, 8558 Japan.
The specific causes of prostate cancer are not known. However, multiple etiologic
factors, including genetic profile, metabolism of steroid hormones, nutrition, chronic
inflammation, family history of prostate cancer, and environmental exposures arethought to play significant roles. Variations in exposure to these risk factors may
explain interindividual differences in prostate cancer risk. Single nucleotide
polymorphism (SNP) arrays provide a high­resolution platform for describing
several types of genetic changes simultaneously. With the resolution of these arrays
increasing exponentially, they are becoming increasingly powerful tools fordescribing the genetic events underlying cancer.
Wehave genotyped morethan 50 non­synonymous missense­SNP ofcancer­related genes between 210 prostate cancer patients and 474 matchedhealthy males in Japan. Eight SNP of 6 genes were significantly affected theincidence of all­age prostate cancer. These genes included 3 DNA­repair genes, 2tumor suppressor genes and so on. Compared between age­groups > 70y and <70y,the repertoire of the risk­associating SNPs were largely different each other (7 SNPof 6 genes VS 10 SNP of 9 genes). Cancer­association of these 13 of 15 SNP is novel
findings. As result, prediction of the Life­Time Risk for prostate cancer in Japanese
Males was done. The highest risk groups exhibited about 33% (mean=1.5%)
life­time risk for >70y onset, and 18% (mean=2.8%) for <70y onset.
Thus, our novel way of combining the risk factors of the statisticallysignificant SNP genotypes would be useful for clinical application to predict
prostate cancer predisposition of males in Japan and may contribute for prevention
and early detection of the disease. This strategy can be applied for othermalignancies and for other ethnic populations by choosing proper SNP
combinations.

62. Cooperative Research on Genetic Predisposition for prostate Cancer in Asia
by Analyzing Cumulative Association of SNPs ­The analyses data in Chinese­
Lei Wang 1), Peng Huang 2), Haruki Kaku 2,3), Ming Li 4), Kai Yang 5), Liping Xie
5),Akiko Sakai 6), Kenji Shimizu 6), Hiromi Kumon 2,3) & Yanqun Na 1,8)

1) Department of Urology, Peking University People’s hospital, Beijing, China.
2) Department of Urology, Okayama University Graduate School of Medicine,
Dentistry and Pharmaceutical Sciences, Okayama, Japan.
3) Center for Gene and Cell Therapy, Okayama University Graduate Schoolof Medicine, Dentistry and Pharmaceutical Sciences, Okayama, Japan.
4) Department of Urology, Peking University School of Oncology, Beijing
Cancer Hospital & Institute, Beijing, China.
5) Department of Urology, the First Affiliated Hospital, School of Medicine,
Zhejiang University, Hangzhou, Zhejiang Province, China.
6) Department of Molecular Genetics, Graduate School of Medicine, Dentistry,
and Pharmaceutical Sciences, Okayama University, Japan.
7) Peking University Wu Jieping Urology Medical Center, Peking University
Shougang Hospital, Beijing, China.
BACKGROUND In a recent study of Okayama University, a list of missense single
nucleotide polymorphisms (SNPs) had been found to be associated with risk of
prostate cancer (PCa) in Japanese population. These SNPs also manifested
distinguished accumulative effects in predicting the risk of PCa. AIM To explore
whether these SNPs had similar effects in Chinese Han population, which are
geographically and genetically close to Japanese population.
PATIENTS AND METHODS One hundred and twenty­five PCa patients and 100
cancer­free controls, which were recruited from four university hospitals in Beijing
and one university hospital in Hangzhou between January 2009 and March 2010,
were included in the cancer association study. Fourteen missense SNPs, eachlocated in different genes, were genotyped using a Snapshot assay.
RESULTS Seven SNPs were found to be significantly associated with risk of PCa.
Among them, 4 SNPs had high­risk associations (Odds Ratio range 1.89­7.84) andthe other 3 had protective effects (Odds Ratio range 0.30­0.61). The seven related
genes included 4 DNA­repair genes and 1 each of tumor suppressor gene, apoptosis
regulator gene and chromosome­segregation gene. Cumulative effects were also
revealed for these 7 SNPs in predicting prostate cancer risk. The life­time risk of7

PCa occurrence for the highest­risk group we appointed was 31.2 times higher thanthat of the lowest­risk group (12.8% vs. 0.41%).
CONCLUSIONS According to this small size, case­control study, many missense
genetic variants might contribute to the genetic predisposition to prostate cancer in
Chinese Han population. These variants also manifested certain effects inpredicting PCa occurrence. Larger studies with sufficient power will be needed toconfirm our findings.

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3. Complications of Urological Laparoscopic Surgery: A Single Institute Experienceof 1,017 Procedures
Takaaki Inoue, Hidefumi Kinoshita, Masahiko Satou, Naoki Oguchi, Gen Kawa,
Kouei Muguruma, Takayuki Murota and Tadashi Matsuda
Department of Urology and Andrology, Kansai Medical University, Osaka Japan
Background and Purpose: We evaluated complications of urological laparoscopic
surgery at our institution.
Patients and Methods: One thousand and seventeen urological laparoscopic
surgical procedures were performed in Kansai Medical University from December
1991 to January 2009, including 277 radical prostatectomies, 13 donor
nephrectomies,

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partialnephrectomies,

pyeloplasties,

97

nephrouretectomies,

54

158

radicalsimplenephrectomies,
nephrectomies,

55
128

adrenalectomies, 34 varicocelectomies, and 127 other procedures. Medical records ofeach procedure were retrospectively evaluated. The difficulty of each procedure wasclassified according to the European Scoring System. Intra­ and postoperativecomplications were graded according to the Satava and Clavien classifications,
respectively.
Results: Among the 1,017 laparoscopic procedures, 148 complications occurred in
123 patients, resulting in a total complication rate of 14.6%. Conversion to open
surgery occurred in 20 patients (1.9%). Nephrouretectomy had the highest incidenceof complications at 23.7%, which was significantly higher than that of otherprocedures classified as Difficult Group (D­Group) according to the European
Scoring System (p<0.05). Clavien grades I and II accounted for 73.8% of all the
postoperative complications. We experienced one fatality caused by air embolism.
Conclusion： We evaluated the complications of each procedure using the European
Scoring System for classification of technical difficulty. Based on the results of ourretrospective study, nephrouretectomy should be upgraded as “very difficult” groupaccording to the European Scoring System. Appropriate grading by technicaldifficulty is beneficial for the prevention of complications from laparoscopic surgery.

94. Percutaneous renal surgery performance on live pigs can be improved bypracticing on a biologic porcine kidney model.
Zhang Yi1)

Wang Gang2),

Na Yanqun1)

1)

Wu Jie Ping Urology Center, Peking University

2)

Urological institute of Peking University

OBJECTIVES To investigate how the learning and training on a biologic porcine
kidney model can improve the performance of percutaneous renal procedure on live
pigs
METHODSA previously reported biologic bench model is set up using a porcine

kidney. Following instruction and demonstration, novice urologists are asked topractice on this model under X­ray guidance first and then perform percutaneous
procedure on live pigs using the same manipulations. The time of successful
puncture, tract establishment and intrarenal exploration on the model and on live
pigs are compared to investigate if a better performance is achieved.
RESULTS

20 novice urologists were investigated using this method. Significant

reduction

of

time

spent

on

successful

puncture(p=0.002)

and

tract

establishment(p=0.018). No difference is observed on the time of intrarenal
exploration.
CONCLUSIONS Training on the biologic porcine kidney model can help trainees
to shorten the crucial step time of percutaneous renal surgery and improve theirperformance on live pigs. This model is simple and easy to build with readily
available materials. It provides realistic and reproducible practice and may beinteroperated as one of invaluable teaching tools for percutaneous renal surgery in
the laboratory.

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5. Primary congenital bladder diverticula in adult.
Wang Jianbo, Sun Xishuang.
Department of Urology The 1st affiliated hospital of Dalian Medical University
Dalian CHINA 116011

Abstract: Congenital bladder diverticula are uncommon, most cases have been
described exclusively in male children, they are very rarely found in adults. We
report on a case of a 43­year­old man who presented with history of urinary
obstruction and recurrent urinary tract infections. Two big congenital bladder
diverticula were diagnosed by physical examinations. Open surgical approaches
were used in this case and a well healed bladder was made.

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6. High­intensity focused ultrasound as salvage therapy for patients with recurrent
prostate cancer after external beam radiation, brachytherapy or proton therapy
Toyoaki Uchida, Sunao Shoji, Mayura Nakano, Satoko Hongo, Masahiro Nitta, andYoshihiro Nagata
Department of Urology, Tokai University Hachioji Hospital, Hachioji, Tokyo,
JAPAN
OBJECTIVES: To investigate the use of high­intensity focused ultrasound (HIFU)
as a salvage therapy in patients with recurrence of localized prostate cancer after
external beam radiation (EBRT), brachytherapy, or proton

therapy.

PATIENTS AND METHODS: We retrospectively reviewed the charts of all
patients who had undergone salvage HIFU for biopsy­proven prostate cancer after
primary radiation therapy. Patient characteristics and oncological outcomes wereassessed.
RESULTS: Records of 22 patients with a median (range) follow­up of 24 (5–80)
months were reviewed. Patients were men with presumed organ­confined disease
who had been treated with salvage HIFU following recurrent disease after EBRT
(fourteen

patients),

brachytherapy(fivepatients:

four

with

high­dosebrachytherapy using In In192 and one with low­dose brachytherapy using Au98) or
proton therapy (three patients). The median (range) age at salvage HIFU was 65(52–80) years, with a median (range) prostate­specific antigen (PSA) level before
radiation therapy of 14.3 (5.7–118) ng/mL and a median (range) PSA level of 4.0(1.2–30.1) ng/mL before HIFU. The median (range) period to HIFU after radiation
therapy was 36 (4–96) months. The biochemical disease­free survival (bDFS) rate inall patients at 5 years was 52%. Rates of bDFS in low­, intermediate­ and high­risk
groups were 100%, 86%, and 14%, respectively. One of the twelve patients who
received post­HIFU prostate biopsy showed malignancy. Side effects included
urethralstricture in four patients, grade I urinary incontinence in four patients,

rectourethral fistula and epididymitis in one of each patient.
CONCLUSIONS: Salvage HIFU is a promising treatment option for local
recurrence after radiation therapy, with morbidity comparable with other forms ofsalvage treatment.

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7. Prognostic factors for renal cell carcinoma with bone metastasis: who are the longsurvivors?
Haruki Kume1), Shigenori Kakutani2), Yukio Yamada3), Mitsuru Shinohara3),
Takashi Tominaga2), Motofumi Suzuki1), Tetsuya Fujimura1), Hiroshi Fukuhara1),
Yutaka Enomoto1), Hiroaki Nishimatsu1), Yukio Homma1)
1). Department of Urology, the University of Tokyo Hospital2). Department of Urology, Mitsui Memorial Hospital3). Department of Urology, Tokyo Metropolitan Cancer and Infectious diseases
Center Komagome Hospital
Purpose:To develop a predictive model for overall survival in renal cell carcinoma(RCC) with bone metastasis.
Patients and method: We retrospectively collected RCC cases with bone metastasis
treated at three tertiary referral centers. Clinicopathological parameters andoutcome data were analyzed to detect the predictors for overall survival.
Results: The sample comprised of 70 males and 26 females with a median age of
59.7 years. Histological diagnosis was clear cell RCC in 64, papillary RCC in 5,
collecting duct RCC in 2, and unclassified cancer in 7. Sarcomatoid differentiation
was found in 18. The metastases were detected synchronously (n=44) or
metachronously (median interval, 32.2 months; n=52). The median overall survival
was 9.25 months. Multivariate analysis identified sarcomatoid differentiation(p=0.012), vertebral bone involvement (p=0.043), extraosseous metastasis (p=0.013),
elevated lactate dehydrogenase (1.5 times upper limit, p=0.003), and elevatedC­reactive protein (>0.3 mg/dl, p=0.002) as the significant risk factor. According tothese risk factors, we reclassified our cases into three groups; low risk (0 ­1 risk
factor, n=24), intermediate risk (2 ­ 3 risk factors, n=55), and high risk (4­ 5 risk
factors, n=17). This grouping clearly separated the survival among these groups (allp<0.001 by log­rank test). The prediction was more accurate than MSKCC
classification system.
Conclusions: Our risk classification incorporating five risk factors provides anaccurate prediction of survival thus would be helpful for clinical decisions for RCC
with bone metastasis.

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8. The expression and clinical value of cytokeratin in peripheral blood of patientswith urinary carcinoma.
Xiaoqiang Liu, Jiang Weifan,Zhang Gaofeng,Sun guang.
Urology Dep. 2nd hospital of Tianjin Medical University,Tianjin Institute of
Urology
Objective: In this article, we present our researches in early diagnosis of
micro­metastasis by detecting the presence of CK in the circulating peripheral blood
supplemented with the detection of CD. Two common urinary carcinomas: renal cell
carcinoma (RCC) and bladder transitional cell carcinoma (BTCC) are included inthis study. To make the presentation clear and for the convenience of discussion, I’dlike to present our studies of RCC and BTCC separately.
First, in the RCC group, when these factors were found, we carried out an analysisof their level in peripheral blood in RCC patients, which was followed by evaluationof the value of CK19 as a marker of CTC.
Methods: We chose 51 patients of renal cell carcinoma (RCC) for this study and 32
healthy volunteers as the control group. We analyzed and recorded the level ofCD45­ CK19+ of the test group according to age, gender, grade, staging and so on.
Follow­up was to get the information of their general condition and location ofmetastasis. All mRCC cases had received targeted therapy of sorafenib. Diametersof all metastasis before and after treatment were measured. Therapeutic effect wasgraded as CR, PR, SD, PD. The results were analyzed . Finally, we used SPSS 13.0
to analyze the level of CK19 of different clinical staging and different grading.
Results:

3 cases of 32 in control group are positive with a rate of CK19 at 9.38%.

37 cases of 51 in the test group are positive, with a rate of CK19 at 68.63%. The
differences of two groups have statistic significance (P<0.01). The numbers of casesof positive and negative in CK19 are 7 and 1 before operation. The numbers of casesof positive and negative CK19 are 5 and 2 after operation. At the end of theexperiment, the numbers of CR、PR、SD、PD and dead are 1、3、8、2 and 1
respectively.

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9. Systemic T­cell activation following in situ gene therapy in prostate cancer
patients
Shinji Kurosaka1), Takefumi Satoh1), Makoto Kubo2), Ken­ichi Tabata1), Kazumasa
Matsumoto1), Fumiya Obata2), Yasutomo Nasu3), Haruki Kaku3), Hiromi Kumon3),
Malcolm K. Brenner4), Timothy C. Thompson5) and Shiro Baba1)
1)Department of Urology Kitasato University School of Medicine

2) Division of Immunology Kitasato University School of Allied Health Sciences
3) Department of Urology Okayama University Graduate School of Medicine
4) Center for Cell and Gene Therapy Baylor College of Medicine
5) Department of Genitourinary Medical Oncology MD Anderson Cancer Center
Introduction and Objectives: Neoadjuvant in situ cytotoxic gene therapy canpotentially trigger a systemic immune response, which could impact occult
micro­metastatic disease. We are currently conducting a clinical trial using in situ
adenoviral vector mediated Herpes Simplex Virus­thymidine kinese (HSV­tk) geneplus ganciclovir (GCV) therapy. This study evaluates the systemic T­cell response
following gene therapy in patients with high­risk prostate cancer.
Methods:

This study was approved by the ethical committee of Kitasato

University, and by the Ministry of Health, Labour and Welfare Japan. Threepatients (Pt1, Pt2, and Pt3) who agreed this Phase I/II trial were repeatedly given 2
weeks of intravenous GCV administration following the intraprostatic HSV­tk
injection. PBMC were collected for 7 days from HSV­tk injection and post treatment.
PBMC were analyzed by flow cytometry. We also measured intracellular IFN­
produced by T cells to responding MHC­I restricted prostate specific antigen (PSA)
peptides. Humoral responses were examined by Western blot.
Results: The activated T cells transiently increased in both the first and second
HSV­tk+GCV treatments. The activated CD8+T cells of Pt2 and Pt3 markedly
increased during second treatment. In Pt1 and Pt2, frequencies of PSA­peptides
specific CD8+T cells slightly increased after second treatment. Although the tumor
antigen was unidentified, we detected the antibody that increased during
treatments in Pt2.
Conclusions: We present evidence of systemic T­cell responses following HSV­tk +GCV gene therapy under clinical trial conditions. There was an increase inactivated CD8+ T cells in the peripheral blood following vector injection suggesting
the potential for activation of components of cell­mediated immune response in trial
conditions.

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10. The expression and role of transient receptor potential channel A1 in the
bladders of rat and human being.
Shuqi Du
Department of Urology, the First Affiliated Hospital, China Medical University,
Shenyang, China

Objectives: Transient receptor potential channel A1 (TRPA1) is a candidate for bothmechanosensory transduction and nociception. We investigated its expression inboth rat and human bladders, and examined its role in micturition.
Methods: Real­time PCR and immunohistochemistry were utilized to examine theexpression level and localization of TRPA1 in both bladder and dorsal root ganglion(DRG). The function of TRPA1 in micturition was analyzed by cystometry in either
normal or capsaicin­pretreated rats. The human bladder specimens were divided
into normal and bladder outlet obstruction (BOO) groups.
Results: The expression level of TRPA1 mRNA in rat DRG/bladder mucosa/bladdermuscular layer was 199:16:1. Of the bladder­innervating DRG neurons, 50.8%
showedTRPA1immunoreactivity.

Intravesicalperfusion

of

600µM

trans­cinnamaldehyde significantly decreased the intercontraction interval andpressure threshold to 59.8% and 83.8% of their control values, respectively. These
effects were reversible by washing out the drug. Desensitization of C­fibers bycapsaicin markedly attenuated the trans­cinnamaldehyde effects. Similarly, 400µM
isothiocyanate reversibly and significantly decreased the intercontraction interval
to 64.7% of its control value.
The expression level of TRPA1 mRNA in human bladder mucosa/muscularlayer/prostate was in the ratio of 639:1:16. TRPA1 mRNA in the bladder mucosa
with BOO was significantly upregulated to 2.32 times of control. TRPA1 protein was
localized in the epithelial cells of both urinary bladder and prostate gland.
Conclusions: TRPA1 is expressed in the bladder­innervating primary sensory
neurons of rat. Its agonists cause bladder hyperreflexia through C­fiber pathway.
TRPA1 is also expressed in the human urinary bladder and prostate. TRPA1 might
be involved in the bladder sensory transduction as a mechanotransducer and/or a
nociceptor. It might also be involved in the induction process of overactive bladderby BOO.

16

Reference:
1. Du S, Araki I, Yoshiyama M, et al. Transient receptor potential channel A1
involved in sensory transduction of rat urinary bladder through C­fiber pathway.
Urology 70: 826­31, 2007.
2. Du S, Araki I, Kobayashi H, et al. Differential expression profile of cold (TRPA1)
and cool (TRPM8) receptors in human urogenital organs. Urology 72: 450­5,
2008.

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11. Synergistic apoptosis of lexatumumab and cisplatin against human renal cell
carcinoma cells in a caspase­dependent manner
Xiu­Xian Wu, Takuma Kato, Hiromi Hirama, Hiroyuki Tsunemori, Xia Zhang,
Motoki Yamashita, Masashi Inui, Mikio Sugimoto, and Yoshiyuki Kakehi
Department of Urology, Faculty of Medicine, Kagawa University, Kagawa, Japan.
1750­1 Ikenobe, Miki­cho, Kita­gun, Kagawa 761­0793, Japan.
Introduction and Objectives: Lexatumumab is a human agonistic antibody against
TNF­related apoptosis­inducing ligand­receptor 2 (TRAIL­R2), which induce
apoptosis specifically in cancer cells. In this study, we investigated whether
chemotherapeutic agents enhance lexatumumab­mediated apoptosis against
human renal cell carcinoma (RCC) cells.
Methods: Cytotoxicity and synergy was assessed by MTT and isobolograpic analyses,
respectively. TRAIL­R2 expression was detected by real­time RT­PCR, flow
cytometry, and western blotting. Apoptosis was assessed by DNA Ladder andcaspase assays.
Results: Treatment of ACHN human RCC cells with cisplatin in combination withlexatumumab had a synergistic cytotoxicity. Synergy was also achieved in six
primary RCC cell cultures. Lexatumumab and cisplatin also synergisticallyenhanced apoptosis. Pretreatment with cisplatin followed by lexatumumab resulted
in high cytotoxicity compare with the reverse sequence. Cisplatin significantly
increased TRAIL­R2 expression at both mRNA and protein levels. Furthermore, thecombination of lexatumumab and cisplatin significantly enhanced caspase­8 activity,
Bid cleavage, up­regulation of Bax, cytochrome c release, and caspase­9, caspase­6,
and caspase­3 activities. Importantly, the activation of capase­8 was significantly
abrogated by the specific inhibitors of caspase­9, caspase­6, and caspase­3.
Furthermore, combination­induced cytotoxicity was significantly suppressed by
DR5:Fc chimeric protein and the specific inhibitors of caspase­8, caspase­9,
caspase­6, and caspase­3. Conclusions: Cisplatin sensitizes RCC cells tolexatumumab­induced apoptosis by potentiation of the extrinsic and intrinsic
apoptotic pathways that lead to enhancement of caspase activation, particularly
caspase­8. These results suggest that combination of cisplatin plus lexatumumab ispromising against RCC.

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