bluuebunny/crossref_metadata_2025_split huggingface dataset:
Prognostic Significance of hOCT1 and ABCG2 Expression in Chronic Myeloid Leukemia Patients Treated with Imatinib
Authors: Kim Yeo-Kyeoung , Kim Hee-Nam , Yu Ri , Lee Il-Kwon , Ahn Jae-Sook , Yang Deok-Hwan , Lee Je-Jung , Kim Hyeoung-Joon 
Published: April 2020

Abstract
Background: Imatinib mesylate is a substrate for both the drug influx transporter human Organic Cation Transporter 1 (hOCT1; SLCA22) and the efflux transporter ATP-binding cassette transporters (ABCG2; BCRP). The correlation between the expression of these two transporters of imatinib and the clinical outcome of imatinib in chronic myeloid leukemia (CML) patients remain clarified.
Methods: hOCT-1 and ABCG2 mRNA levels were determined by performing real-time polymerase chain reaction assays on 83 BM samples obtained at initial diagnosis from the patients with chronic phase CML treated with imatinib (started at 400mg/day).
Results: Of patients with higher hOCT-1 mRNA expression than median value (≥ 34.53 a.u.) at diagnosis, 59.5% achieved complete cytogenetic response (CCyR) at 6 months, whereas of patients with no more than a median hOCT-1 expression, 33.3% could achieve CCyR at 6 months (p=0.035). Furthermore, patients with low hOCT-1 showed higher rate of suboptimal response [failed to achieve complete hematologic response (CHR) by 3 months, major cytogenetic response (MCyR) by 6 months, CCyR by 12 months, and major molecular response (MMR) by 18 months] than those with high hOCT-1 expression (55.9% vs. 33.3%. p=0.048). There was no statistical significance in progression-free survival (PFS) or overall survival (OS) according to the hOCT-1 expression levels. Patients with low ABCG2 expression (&lt; 13.53 a.u.) showed higher MMR rate at 18 months (57.1% vs. 35.5 %, p=0.09) and longer PFS (3 yr PFS, 87.2% vs. 64.1%, p=0.07) than those with high ABCG2 expression but there was no statistical significance. The time to achieving MMR was significantly shorter in patients with low ABCG2 expression than in those with high ABCG2 expression (6.9 ± 2.1 ms vs. 12.2 ± 4.2 ms, p=0.09). The patients with high hOCT-1 expression showed shorter time to achieving MMR, but there was no statistical significance (8.1 ± 1.9 vs. 10.5 ± 1.7, p=0.50).
Conclusions: This study demonstrates that pretreatment assay of hOCT-1 and ABCG2 expression may help us to identify the treatment outcome in the CML patients treated with imatinib.
Modification of IGF1R Expression on Malignant Lymphoid Cells by a Proteasome Inhibitor
Authors: Terui Yasuhito , Mishima Yuji , Mishima Yuko , Hatake Kiyohiko 
Published: April 2020

Abstract
Purpose: The type 1 IGF receptor (IGF1R) is a transmembrane tyrosine kinase, which is overexpressed by tumors, and mediates proliferation and apoptosis protection for tumor cells. IGF signaling affects hypoxia signaling, protease secretion, tumor cell motility, and adhesion. ProIGF1R is cleaved into alpha and beta chains by processing, and their heterodyne moves onto plasma membrane. Folding after transfer to Golgi apparatus modifies the beta chain, and it is also known to degradate by ubiquitin-proteasome system. Bortezomib is a proteasome inhibitor for multiple myeloma, and expression of chaperon proteins such as IGF1R could be increased by bortezomib. Here, we examined and analyzed expression of IGF1R on malignant lymphoid cells, and discuss possibility of IGF1R as a target in the treatment of lymphoid malignancies.
Methods: IGF1R expression of malignant T cell lymphoid cell line Jurkat, myeloma cell line IM-9, and bortezomibresistant IM-9 was examined by flow cytometric and western blot analyses. The cell lines were cultured with IGF1 and IGF2 with or without bortezomib, and the growth and cell death were observed.
Results: The IGF1R expression on Jurkat cells was weak by flow cytometry, and western blot analysis revealed that IGF1R beta expression was quite low in spite of higher expression of ProIGF1R and IGF1R alpha. The IGF1R expression on bortezomib-resistant cells was stronger than that on parent IM-9 cells by flow cytometric and western blot analyses. While IGF1 and IGF2 did not give an advantage for the growth of lymphoid cells significantly without bortezomib, there was an advantage for survival of them under exposure to bortezomib.
Conclusion: Difference of IGF1R expression was observed among several malignant lymphoid cells. Cell surface modification by drugs such as bortezomib will make lymphoid cells increased IGF1R expression, and IGF1R inhibition will be a promising way for the treatment of malignant lymphoid cells.
Daunorubicin Loaded Magnetic Nanoparticles of Fe3O4 Greatly Enhance the Responses to Chemotherapy and Induce Apoptosis of Multidrug-Resistant K562 Cells in a Human-Nude Mice Xenograft Leukemia Model
Authors: Chen Bao-An , Lai Bin-bin , Cheng Jian , Gao Feng , Xu Wen-lin , Shen Hui-lin , Ding Jia-hua , Gao Chong , Sun Yun-yu , Wang Jun , Zhao Gang , Song Hui-hui , Bao Wen , Sun Xinchen , Cheng Hong-yan , Deng Yu-xia , Li Guo-hong , Liu Li-jie , Chen Wen-ji , Shao Ze-ye , Xia Guo-hua , Wang Jue-qiong , Yuan Peng , Pei Xiao-ping , Chen Ning-na , Li Qin-ning , Wang Xue-mei 
Published: April 2020

Abstract
Object: To the effect of Fe3O4-magnetic nanoparticle loaded with DNR on multidrugresistant K562 cells in vivo.
Methods: K562-n and its MDR counterpart K562-n/VCR cell were inoculated subcutaneously into both sides of the back of nude mice (5×106 cells/each) to establish a human leukemia xenograft model. The mice were randomly divided into group A receiving normal saline every other day for 20days, group B receiving DNR every other day for 20days, group C receiving Fe3O4-magnetic nanoparticle every other day for 20days, group D receiving Fe3O4-magnetic nanoparticle loaded with DNR every other day for 20days, and group E receiving Fe3O4-magnetic nanoparticle containing DNR every other day for 20days with a magnetic field built on the surface of the tumor tissue. The tumor volume was measured on the day 1, 5, 9, 13, 17 and 21d after the first treatment. Tumor tissues were isolated for examination of the expression of mdr-1, bcl-2, bax and caspase-3 by reverse transcription polymerase chain reaction and Western blotting.
Results: For K562-n/VCR tumor, the tumor volume was markedly lower in groups D and E than in groups A, B and C (group D or E vs group A, B or C, P &lt; 0.05). The transcription of mdr-1 and Bcl-2 gene was significantly lower in groups D and E than in groups A, B and C (group D or E vs group A, B or C, P &lt; 0.05). So did the protein expression of Bcl-2. However, there were no differences among these groups about the protein expression of P-gp. The protein and mRNA expressions of Bax and Caspase-3 in groups D and E were increased significantly compared with groups A, B and C (group D or E vs group A, B or C, P &lt; 0.05). The tumor volume of K562-n was markedly lower in groups C, D and E than in groups A and B (group C, D or E vs group A or B, P &lt; 0.05).
Conclusion: In conclusion, DNR loaded Fe3O4-magnetic nanoparticles can suppress the growth and induce apoptosis further on the MDR K562-n/VCR tumor in vivo compared to DNR alone but not on the K562-n tumor. The external magnetic field failed to improve the antitumor effect.
Pharmacogenetic Outcomes of Pediatric Acute Lymphoblastic Leukemia in Korea
Authors: Kang Hyoung Jin , Kim Hyery , Seo Young Jin , Jang Mi Kyung , Oh Yongtaek , Han Byoung-Don , Park Kyung Duk , Kim Chul Woo , Shin Hee Young , Ahn Hyo Seop 
Published: April 2020

Abstract
Treatment result of pediatric acute lymphoblastic leukemia (ALL) has been markedly improved, but treatment related toxicities and relapse are still remaining problems. Genetic polymorphism is an important factor in the effectiveness and toxicity of anti-leukemic drugs and pharmacogenetics are beginning to emerge as useful research filed to solve those problems. In our experience in the treatment of ALL, many Korean patients could not tolerate full dosages of Western protocols. To make basis for individualized therapy with pharmacogenetics, we analyzed major genes implicated in the treatment of ALL. Fourteen genes of total 103 patients with ALL were analyzed with TotalPlex gene amplification methods (Mol Cell Probes.2008. 22: 193). Among the drug related genes (percentage of mutant type) including CYP3A4*1B (0%), CYP3A5*3 (0%), GSTP1 (22.3%), GSTM1 (20.4%), GSTT1 (16.5%), MDR1 exon 21 (76.3%), MDR1 exon 26 (61.2%), MTHFR (64.1%), MTHFR 1298 (29.1%), NR3C1 1088 (0%), RFC 80 (79.6%), TPMT combined genotype (7%), VDR intron 8 (10.7%), VDR FokI (67%), incidence of mutant was higher in GSTM1 deletion, lower in MTHFR 1298, TYMS enhancer repeat, and VDR intron 8 comparing with the data of Western whites (Rocha J.C. et al. Blood 2005). As we had modified the dose of anti-leukemic agents depending on the toxicity during the treatment, we analyzed the relationship between the dose percent of actually administered dose and the distribution of each mutant to find out polymorphisms affecting toxicities of chemotherapeutic drugs. The mean dose percent of mercaptopurine was lower in patients with variant TPMT then those with wild type (33.2% vs. 53.5%, P=0.04), but there was no polymorphism that influenced the dose percent of methotrexate, daunorubicin, doxorubicin, and L-asparaginase. There was no significant difference in the incidence of genotypes between risk groups and individual mutant did not affect long term survival and relapse in Korean patients with ALL. In conclusion, we found some difference in the incidence of mutant genotypes related to the pharmacogenetics of ALL between Korean and Western whites, but there was no individual genetic polymorphism that affect on the treatment outcome. We expect more extensive researches about pharmacogenetics of Korean to establish the basis for individualized therapy with the consideration of ethnical difference.
Mechanism Research of Reversal of Multidrug Resistance by the Application of 5-Bromotetrandrine and Magnetic Nanoparticle of Fe3O4 Combined with Daunorubicin in a Human-Nude Mice Xenograft Model
Authors: Chen Bao-An , Wu Ya-nan , Cheng Jian , Gao Feng , Xu Wen-lin , Shen Hui-lin , Ding Jia-hua , Gao Chong , Sun Yun-Yu , Wang Jun , Zhao Gang , Song Hui-hui , Bao Wen , Sun Xin-chen , Cheng Hong-yan , Deng Yu-xia , Li Guo-hong , Liu Li-jie , Chen Wen-ji , Wang Jue-qiong , Yuan Peng , Pei Xiao-ping , Chen Ning-na , Wang Xue-mei 
Published: April 2020

Abstract
Objective: To establish the xenograft leukemia model with stable multiple drug resistance in nude mice; to investigate the reversal effect of 5-Bromotetrandrine and Magnetic nanoparticle of Fe3O4 combined with DNR in vivo and to search for the possible reversal mechanisms.
Methods: K562 and K562/A02 cells were respectively inoculated subcutaneously into back of athymic nude mice (1×107 cells/each) to establish the xenograft models. The tumor formation was evaluated by animal ultrasonic inspection. Tumors-bearing nude mice were assigned randomly to five groups which were treated with NS (A group); DNR 1mg/kg (B group); nanoparticle of Fe3O4 combined with DNR 0.63mg/kg(C group): 5-BrTet 2.5mg/kg combined with DNR(D group); 5-Bromotetrandrine 2.5mg/kg and Magnetic nanoparticle of Fe3O4 combined with DNR 0.63mg/kg(E group) respectively. The incidence of tumor formation, growth characteristics, weight and volume of tumor were observed. The histopathologic examination of tumors and organs were detected. For resistant tumors, the protein levels of P-glycoprotein (P-gp) were detected by Western blot.
Results: The tumor incidence was 100% in the nude mice inoculated with either K562 or K562/A02 cells. In 6 to 9 days,the tumors reached a volume of more than 1 00 mm3. In vivo, MTT assay showed K562/A02 tumor maintained the drug resistance. For K562 cells xenograft tumors, there were no apparent differences in tumor suppression effect between the B AC AD AE group. For K562/A02 cells xenograft tumors, 5-BrTet and Magnetic nanoparticle of Fe3O4 combined with DNR significantly suppressed growth of tumor: the inhibition rate was 62.76% while DNR alone be used, the inhibition rate was 3.68%. Pathologic examination of resistant tumors showed the tumors necrosis obviously in E group. Application of 5-BrTet and Magnetic nanoparticle of Fe3O4 inhibited the overexpression of P-gp.
Conclusion: The xenograft leukemia nude mice model was maintain the multiple drug resistance. 5-Bromotetrandrine and Magnetic nanoparticle of Fe3O4 combined with DNR had a significant tumor-suppressing effect on MDR leukemia cells xenograft model.
Effect of Tetrandrine and Toremifene on the Reversion of Multidrug Resistance of K562/A02 Cell Line
Authors: Chen Bao-An , Zhao Qiu-xia , Cheng Jian , Gao Feng , Xu Wen-lin , Shen Hui-lin , Ding Jia-hua , Gao Chong , Sun Yun-Yu , Wang Jun , Zhao Gang , Song Hui-hui , Bao Wen , Sun Xin-chen , Cheng Hong-yan , Deng Yu-xia , Li Guo-hong , Liu Li-jie , Chen Wen-ji , Wang Jue-qiong , Yuan Peng , Pei Xiao-ping , Chen Ning-na , Wang Xue-mei 
Published: April 2020

Abstract
Objective: To study the reversal effect of Tetrandrine(Tet) and the estrogen-receptor inhibitor,toremifene(Tor),on multidrug resistance cell line K562/A02 and to investigate the reversal mechanism of this combination.
Methods: ADM accumulation and the apoptosis percentage of K562 and K562/A02 cells were analyzed by fluorospectrophotometry, respectively. The protein levels of P-glycoprotein (P-gp) were detected by fluorospectrophotometry. The mRNA levels of mdr1 and Survivin were determined by RT-PCR.
Results: The IC50 of ADM for K562/A02 and K562 cells were 57.43±4.55mg/L and 1.16±0.05mg/L respectively. Pretreating K562/A02 cells with toremifene(2.5μmol/L) or Tet(1μmol/L) for 72 hours partially restored the sensitivity of K562/A02 cells to ADM (IC50 were 20.74±1.62mg/L and 14.12±1.20mg/L respectively) but had no effect on K562 cells; IC50 of combined tetrandrine and toremifene was 9.14±1.03mg/L;K562/A02 showed apoptotic characteristics after treated with tetrandrine, toremifene (alone or combination);tetrandrine and toremifene (alone or combination) elevated the intracellular ADM accumulation in K562/A02; P-gp, mdr1 and Survivin mRNA were down regulated.
Conclusion:
Tetrandrine, toremifene (alone or combination) showed significant MDR reversal activity in vitro The reversal activity may be related to the inhibition of P-gp overexpression and down regulation the expression of mdr1 and Survivin mRNA to increase the intracellular accumulation of anticancer drugs, which lead to more K562/A02 cells apoptosis; Multidrug resistance (MDR) can be partially reversed by Tet or Tor of which the combination shows a great synergistic reversal effect.
Variation of p21, P-Gp Expression in K562/A02 Cell Line with Tetrandrine
Authors: Chen Bao-An , Li Jing , Cheng Jian , Gao Feng , Xu Wen-lin , Shen Hui-lin , Ding Jia-hua , Gao Chong , Sun Yun-Yu , Wang Jun , Zhao Gang , Song Hui-hui , Bao Wen , Sun Xin-chen , Cheng Hong-yan , Deng Yu-xia , Li Guo-hong , Liu Li-jie , Chen Wen-ji , Wang Jue-qiong , Yuan Peng , Pei Xiao-ping , Chen Ning-na , Wang Xue-mei , Zhu Min-sheng 
Published: April 2020

Abstract
Object: To study the variation of p21, P-gp expression in reversion of multidrug resistance of K562/A02 leukemic line with different concentration of tetrandrine(TET), to provide new theoretic evidence for the clinical application of TET.
Methods: The IC50 (the concentration causing 50% inhibition of cell growth) of DNR was analyzed by MTT method; The Expressions of p21 were assayed by westernblot; The Expressions of P-gp were assayed by FCM; The expressions of mdr1 were assayed by RT-PCR; The variation of p21 was accentuation with the accrescence concentration of TET(P &lt;0.05).
Results: The variation of p21 protein in K562/A02 cells was accentuation with the accrescence concentration of TET(p&lt;0.05), Mdr1 mRNA was lowly displayed in K562 cells and highly displayed in K562/A02 cells(p&lt;0.01), the variation of mdr1 was attenuation with the accrescence concentration of TET(p&lt;0.05); P-gp was lowly displayed in K562 cells and highly displayed in K562/A02 cells(p&lt;0.01), the variation of P-gp was attenuation with the accrescence concentration of TET(p&lt;0.05).
Conclusion: TET may reverse multidrug resistance of K562/A02 cells by the regulation of p21,P-gp and mdr1.
Factors Involved in Response to Thalidomide in Myelodysplastic Syndrome: Impace of HLA and Prognostic Factors
Authors: Bakanay Sule Mine , Dalva Klara , Topcuoglu Pervin , Civriz-Bozdag Sinem , Arat Mutlu , Beksac Meral , Gurman Gunhan 
Published: April 2020

Abstract
Myelodysplastic Syndrome (MDS) is a clonal disease of hematopoiesis characterized by dysplasia in one or more lineages. Since the syndrome is clinically heterogenous, treatment options are complex and hard to define. Thalidomide (T) is the first agent tested in MDS for its immunomodulatory and anti-angiogenic effects. Twenty to 59 % response rates, which is mainly hematological improvement, have been reported. However, because of excessive toxicity and low response rates, thalidomide has not been widely used as an effective treatment option for MDS. Factors invoved in response to T are not well delineated. Better response to immunosupression (IS) in HLA-DRB1*15(+) patients with aplastic anemia and some MDS subgroups have been reported. In this study, the medical records of MDS patients, who received T at least for ≥ 3 months were retrospectively analyzed. Responses could be evaluated in 47 patients with a median age of 59 years (range, 15–84) and M/F: 29/18. According to WHO classification, there were 7 refractory anemia (RA), 9 refractory anemia with ring sideroblasts (RARS), 5 refractory cytopenia with multilineage dysplasia (RCMD), 2 RCMD-RA, 13 RCMD-RS, 6 Hypoplastic MDS, 1 RAEB, 4 MDS/MF. International Prognostic Scoring (IPSS) risk groups consisted of 42 low/intermediate-1 and 5 intermediate-2/high risk patients. Clonal cytogenetic abnormalities were found in 16 patients (5 trisomy 8, 3 monosomy 7, 2 delY, 2 increased chromosomal breaks, 1 del5q, 1 monosomy 15 and 1 del21q). Thalidomide was administered at a dose range of 100–400mg/day and for a range of 3–48 months. Twenty-one patients had hematological response (10 major and 11 minor). Seven patients among responders had received additional erythropoietin (EPO) combined with T. There was no statistically significant difference between responders and non-responders in terms of age (median 53 vs 60.5 years), sex (M/F: 12/9 vs 17/26) and IPSS risk groups (Low risk/high risk: 19/2 vs 23/3). Response rates were 44% in RA; 50% in RCMD-RA; 56% in RARS; 55% in RCMD-RS; 20% in RCMD; 20% in hypoplastic MDS; 75% in MDS/MF. and none in RAEB patient. The EPO levels did not differ significantly between the responders and non-responders (EPO miu/ml &lt;500 in 93% vs 83%). However, compared with non-responders, majority of the responders had ferritin levels &lt;1000 mcg/L (83% vs 50%, p&lt;0.05). A total of 21 patients had their HLA type screened. HLADRB1*15 was detected in 44% of responders (4/9) and 16% of non-responders (2/12) (p&lt;0.05). Interestingly, among 5 patients who had trisomy 8, 4 responded to T. Fourty-five percent of mainly low risk-MDS patients responded to T. Patients with MDS/MF and RARS/RCMD-RS and patients with less iron overload responded better. HLADRB1*15 predominance and the presence of trisomy 8 in T responders should be underlined.
Prognostic Relevance of Retrospective WHO Re-Classification in 650 MDS Patients Diagnosed by FAB Criteria
Authors: Breccia Massimo , Latagliata Roberto , Cannella Laura , Carmosino Ida , Stefanizzi Caterina , Santopietro Michelina , Alimena Giuliana 
Published: April 2020

Abstract
We retrospectively re-classified according to WHO criteria our series of consecutive 650 MDS cases, in order to validate the prognostic role of this classification in a unicentric series of patients. Based on FAB criteria, 196 patients had been diagnosed as RA, 41 as RARS, 242 as RAEB, 74 as RAEB-t and 83 as CMML. The new WHO classification could be applied to 479 of the 650 patients; of 196 FAB-RA only 76 were classified as “pure” RA, with only anemia and erythroid dysplasia, whereas 87 patients were re-evaluated as RCMD, for the presence of peripheral cytopenias and dysplasia in ≥ 10% of 2 cell lines. Twenty-nine patients had cytopenias but with only unilineage dysplasia in granulocytes or megakaryocytes, and were re-interpreted as MDS, unclassified (MDS-U). Of 41 FAB-RARS, 32 maintained their diagnosis, whereas 9 patients were classified as RCMD-RS, based on criteria listed above. One-hundred-one patients were identified as RAEB-1, for the presence of 5–9% blasts in bone marrow, and &lt; 5% peripheral blasts, whereas 141 patients were classified as RAEB-2 for the evidence of 10–19% and of 5–19% blasts in bone marrow and peripheral blood respectively. We tested the WHO classification in univariate analysis with respect to several clinical features at presentation. Significant correlations were found as to hemorrhagic symptoms in RCMD and RAEB-2 patients (p=0.014), acute transformation, with higher frequency in RCMD and RAEB-2 categories (p=0.002), transfusion dependence, with higher requirement in RAEB-2 (107/141 RAEB-2 vs 59/101 RAEB-1) and in RCMD (63/87 patients) categories; also the application of different scoring prognostic systems, such as the Bournemouth and Spanish, appeared to possess stastical significance (p=0.001 and p=0.002, respectively). No differences were found as to sex, age, infection occurrence and cytogenetic abnormalities among various subgroups. Kaplan-Meier survival test showed high significance, with median overall survival ranging from 55.8 months for pure RA to 22.6 months for RAEB-2 category (p=0.00001). In conclusion, the retrospective application of WHO classification to our series of MDS patients clearly identified prognostic correlations in various disease subtypes. Categorization of MDS entities according to WHO criteria may distinguish parameters of prognostic importance in large groups of patients with morphological similar features.
ABCG2 Expression and Methylated Regulation in Human Multiple Myeloma Cell Lines
Authors: Qu Shiqiang , Fu Chengcheng , Yao Li , Chen Guanghua , Cen Jiannong , Qi Xiaofei , Depei Wu 
Published: April 2020

Abstract
Objective: The expression and regulation of multidrug resistance protein ABCG2 in multiple myeloma (MM) is unclear, expression of ABCG2 mRNA and protein, promoter methylation regulation of ABCG2 in MM cell lines will be investigated.
Methods After human MM cell lines 8226 AU266 and XG-7 were incubated in the demethylation agent 5’-AzaDC, the change of ABCG2 mRNA and protein expression were evaluated separately by using the Taqman probe-based real-time quantitative polymerase chain reaction(RQ-RT-PCR) and flow cytometry. The percentage of methylated alleles was assayed by SYBR green based methylation-specific quantitative PCR (MSQ-PCR). The melting curve was then analyzed to judge the specificity of the PCR product.
Results The three MM cell lines have different expression level of ABCG2 mRNA(Table 1) and protein but all much higher than normal PBMCs and in different promoter methylation status. The highest level of ABCG2 mRNA and protein were detected in the 8266 cell lines and the promotor region is in unmethylation status. Expression in the U266 cell lines was moderate and the methylated percentage is 61.55%±1.98%. Expression in the XG-7 is the least, the methylated percentage is 78.40%±2.46%. ABCG2 expression increased after exposure to demethylation agent 5’-AzaDC to 2 fold in U266 and 3.5 fold in XG-7 cell lines. After MSQ-PCR, the melting curve was analyzed. The Tm value of methylated product and the unmethylated product were 77°C and 75.5°C separately.
Conclution: ABCG2 mRNA and protein expression in 3 MM cell lines were much higher than that in normal peripheral blood mononuclear cells. Demethylation of the promotor increased ABCG2 mRNA and protein expression. So expression of ABCG2 is partially regulated by promoter methylation in myeloma cell lines. The cell line with the more ABCG2 expression may have more potential to effluence the chemotherapy agents. SYBR green I based fluorescence quantitative PCR can calculate the percentage of the methylated alleles in genome.