bluuebunny/crossref_metadata_2025_split huggingface dataset:
Mechanism of inhibitory effect of terpenoid compound from Achillea millefolium on human lung cancer cell proliferation.
Authors: Wang Siming , Wang Yu , Wu Yibing , Dong Mei , Ni Zhiyu 
Published: June 2021

e15058 Background: A large number of reports have found that many components in traditional Chinese medicine preparations have obvious anti-tumor activities, and most of these effective components are realized by inducing tumor cell apoptosis. In this part of the experiment, we studied terpenoid compound (3β-acetoxy-1β,4α-dihydroxy-11αH-eudesman-12,6α-olide Achillea millefolium A) from Achillea millefolium anti-cancer mechanism, to determine whether this compound by inducing cell apoptosis plays its antitumor activity of cell proliferation. Methods: (1) The apoptosis of NCI-H292 cells treated with 1 μmol/L and 10 μmol/L Achillea millefolium A was detected by Annexinov-FITC /PI stain. (2) The apoptosis of NCI-H292 cells treated with 10 μmol/L Achillea millefolium A for 12 h and 24 h was detected by in situ end labeling. (3) The changes of p53, Bax, Caspase-3/8/12 and GRP78 in NCI-H292 cells were detected by Western blotting. Results: (1) Effects of Achillea millefolium A on apoptosis of NCI-H292 cells: The apoptosis rate of NCI-H292 cells treated with 1 μmol/L and 10 μmol/ L Achillea millefolium A for 12 h was (15.31)% and (17.84)% respectively. The apoptosis rate of NCI-H292 cells treated with 1 μmol/L and 10 μmol/ L Achillea millefolium A for 12 h was (15.37)% . (2), TUNEL method further verified the cell apoptosis: NCI-H292 cells were treated with 10 μmol/L Achillea millefolium A for 12 h and 24 h, and the positive cells accounted for (13.89)% and (28.37)% respectively, compared with blank control group (4.17)%. (3), Achillea millefolium A up-regulated the expression of p53 protein, Bax protein, Caspase-3/8/12 and GRP78 protein in NCI-H292 cells: After NCI-H292 cells were treated with 1 μmol/L and 10 μmol/ L Achillea millefolium A for 24 h, the expressions of p53, Bax, Caspase-3 and Caspase-8 in NCI-H292 cells were significantly up-regulated. After treatment with 1 μmol/L and 10 μmol/ L Achillea millefolium A for 12 h, the expression of Caspase-12 and GRP78 increased significantly. Conclusions: Achillea millefolium A is a terpenoid compound with obvious apoptosis-inducing activity on human lung cancer cell line NCI-H292. Achillea millefolium A induced apoptosis of NCI-H292 cells was the result of multiple pathways. Achillea millefolium A can up-regulate p53 protein, and then up-regulate Bax, and then activate Caspase-3 protein expression. At the same time, the expression of caspase-3 protein was further increased by activating caspase-8, and apoptosis was induced by both exogenous and endogenous pathways. Notably, the complex of ER stress pathway GRP78 and caspase-12 may also be one of the mechanisms by which Achillea millefolium A induces apoptosis in NCI-H292 cells.
Frequency of homologous recombination deficiency gene mutations in melanoma and its relevance to the immunotherapeutic response.
Authors: Liu Huanhuan , Zhang Yanrui , Ding Feng , Zhang Yun , Liang Xiayuan , Lou Feng , Cao Shanbo , Wang Huina 
Published: June 2021

e15073 Background: Homologous recombination deficiency (HRD) can regulate the tumor immune microenvironment based on increasing tumor-infiltrating lymphocytes and therefore might serve as a biomarker of immunotherapeutic response. The HRD phenotype has been defined as the presence of a non-silent somatic mutation in homologous recombination-related (HRR) genes. Here, we focused on frequency of HRR genes mutation and the relationship between HRR genes mutation and clinical immunotherapy responses in melanoma. Methods: We prospectively sequenced 130 Chinese melanoma patients using next-generation sequencing techniques with Acornmed panel with 808 cancer-related genes. 208 patients with WES data from the TCGA project were included. Correlations between HRR gene mutations and clinical outcomes were identified via bioinformatic analysis using TCGA datasets. Results: In the Chinese cohort, 40% (52/130) patients exhibited genomic alterations in HRR genes. The frequently mutated genes were ATM (14%), BRCA2 (13%), CHEK2 (6%), PALB2 (6%), BRIP1 (4.6%), BARD1 (3%), BRCA1 (2%), CDK12 (2%), CHEK1 (1%), FANCL (1%), and RAD54L (1%). In TCGA cohort, 26% (53/208) patients had at least one somatic mutation in an HRR gene. The frequently mutated genes were BRCA2 (9%), BRCA1 (7%), ATM (6%), BRIP1 (4%), PALB2 (4%), BRAD1 (1%), CHEK2 (1%), PPP2R2A (1%), RAD51B (1%), RAD51C (1%), RAD54L (1%), CHEK1 (1%), and RAD51D. Chinese melanoma patients had higher frequently mutated HRR genes compared to that in TCGA patients (40% vs 26%, p=0.005). In addition, compared with TCGA cohort, the Chinese cohort had higher frequency for ATM (14% vs 6%, p=0.019) and CHEK2 (6% vs 2%, p=0.006). The TCGA cohort had a higher frequency of BRCA1 mutations than the Chinese cohort, although the difference was not statistically significant. Unfortunately, we did not find a significant correlation between HRR mutation and immune cells. Interestingly, in the TCGA cohort, patients with HRR-mut had significantly better OS than patients with HRR-wt (log-rank p=0.027). Conclusions: Our data suggest that detection of somatic mutations in HRR genes might contribute to identify patients who might benefit from immune checkpoint blockade therapy. In addition, the molecular features of HRD provide new opportunities to predict the tumor response to multiple treatments.[Table: see text]
Gamma-interferon inducible lysosomal thiolreductase (GILT) effect on breast tumor microenvironment through regulating CXCR6/CXCL16 axis.
Authors: Fei YinJiao , Liang MingXing , Li Lei , Song Yuxin , Liu Zhen , Tang Jinhai 
Published: June 2021

e15081 Background: Gamma-interferon Inducible Lysosomal Thiolreductase (GILT) is constitutively expressed in most antigens endocytosed by antigen presenting cells (APCs), and its function is to catalyze the reduction of disulfide (S-S) bonds in protein substrates. The cytokine CXCL16 is one of the only two known plasma membrane chemokines which induces chemotaxis of activated T cells and bone marrow plasma cells in tumor microenvironment. It contains a free end folded by two sulfur bonds and therefore could also be a zymolyte of GILT. Previous studies found that specific receptor of CXCL16, CXCR6, was significantly overexpressed in breast cancer tissues and metastatic axillary lymph nodes. We suppose whether CXCR6/CXCL16 axis is regulated by GILT and affect tumor microenvironment, thereby eliciting specific anti-tumor immune responses in breast cancer (BC). Methods: GILT expression in BC was evaluated using publicly available data from The Cancer Genome Atlas (TCGA). GILT gene was analyzed in UALCAN ( http://ualcan.path.uab.edu/analysis-prot.html ) . In vitro, Immunohistochemistry (IHC) was conducted to examine the location and relation of GILT and CXCR6. Gene Set Enrichment Analysis (GSEA) was performed to mine the biological pathways involved in BC related GILT regulatory network. The expression of GILT at protein and RNA levels were detected by Western Blot and RT-PCR assay. Overexpression and knockdown of GILT in BC cell lines was carried out to further analyzed the correlation between GILT and CXCL16/CXCR6. Results: TCGA database showed that GILT expression was increased in the stroma of BC compared with normal, and was correlated to shorter BC overall survival. GSEA suggested that the expression of GILT was associated with chemotactic factors. Pearson analysis and IHC showed GILT had a strong correlation with CXCL16/CXCR6 axis in the aspect of angiogenesis and immunity. qRT-PCR and Western Blot assay also revealed that GILT had high expression in BC. Besides, patients with high expression of GILT in IHC simultaneously showed high immunoreactive to macrophage markers, which was related to neovascularization and anti-tumor immune responses. Compared with the normal breast cell line MCF-10A, GILT protein had high expression in Hs578T and low expression in MDA-MB-231 cell line. GILT was overexpressed in MDA-MB-231 and knockdown in Hs578T. Result showed that high level GILT promoted the production of CXCL16/CXCR6,while GILT siRNA knockdown inhibited the production of CXCL16/CXCR6. Conclusions: GILT could catalyze CXCL16 in BC, function as a key mechanism to affect tumor microenvironment through CXCR6/CXCL16 pathway. GILT-activated CXCL16 levels showed a strong connection with unfavorable outcomes in BC, which could be a potential biomarker of prognosis and a novel therapeutic target.
Clinical application of next-generation sequencing in cancer patients.
Authors: Shamrikova Valentina , Shilo Polina , Stepanova Maria , Dekhanova Ksenia , Ledin Evgeny 
Published: June 2021

e15090 Background: Next-generation sequencing (NGS) has been moving rapidly from pure cancer genomics research towards clinical oncology to provide a rigorous personalized approach to treating cancer. Still there is a number of limitations to overcome before we are ready to fully introduce NGS into everyday clinical decision-making. Here we are sharing our experience with using targeted NGS platforms in clinical practice. Methods: We retrospectively evaluated the NGS results and the treatments, provided to the patients after NGS testing between Marсh 2019 and January 2021. The patients’ clinical characteristics, response to treatment and genomic mutation profiles were reviewed. Coprimary endpoints were the percentage of patients with NGS revealed targeted therapy options and the percentage of patients who received mutation-driven therapy. Results: Samples from 95 patients were tested, most frequently GI tumors (n=25, 26%), hepatobiliary tumors (n=11, 12%), breast cancer (n=10, 11%) gynecological tumors (n=10, 11%), central nervous system tumors (n=7, 7%) and others (n=32, 33%). An average of two gene alterations were identified per patient (range 0-15). The 95% (n=90) of samples had at least one detected mutation, and in 42% (n=40) of cases gene mutations happened to be targetable with existing drugs, including off-label prescriptions. Mutation-driven therapy was performed in 9,5% cases (n=10) with clinically meaningful response in 5 patients (2 patients with ovarian cancer, 2 patients with colon cancer and 1 melanoma). Conclusion: Mutational profiling using a targeted NGS panel has identified potentially targetable alterations in a vast majority of advanced cancer patients. The assay has revealed additional therapeutic options in 42% of patients. However, due to the short-term follow-up, just a small number of patients have received mutation-driven therapy. Further study will be required to determine if the mutation-driven therapy will prove to be clinically meaningful in these patients.
Application of a colorectal cancer ctDNA methylation test in endocrine cancers.
Authors: Winter Jean M. , Sheehan-Hennessy Lorraine , Pedersen Susanne Kartin , Wassie Molla M. , Young Graeme P. , Symonds Erin L. 
Published: June 2021

e15533 Background: Assays for circulating tumor DNA (ctDNA) have shown utility for cancer detection and management. It is important to demonstrate cancer specificity of targeted ctDNA biomarkers. Detection of methylated BCAT1 and IKZF1 ctDNA has shown high sensitivity for colorectal cancer (CRC). The aim of this study was to investigate whether hypermethylation of these two biomarkers is also present in prostate or breast adenocarcinoma (PCa or BCa) tissues, and if they are detectable as ctDNA from such cancer patients. Methods: A subset of TCGA data comprising matched tumor and normal samples from PRAD (n = 50), BRCA (n = 90) and COADREAD (n = 44) cohorts were accessed to interrogate DNA methylation (450K) at CpG probes covered by the BCAT1/ IKZF1 ctDNA assay (cg10764357 and cg07589773/cg18607529, respectively). Linear regression of BCAT1/ IKZF1 methylation and tumor staging was performed for cases with AJCC staging data (PRAD n = 503; BRCA n = 788; COADREAD n = 284). Blood was collected from BCa (n = 32) or PCa (n = 101) patients prior to starting treatment at Flinders Medical Centre (South Australia). DNA isolated from plasma was bisulphite-converted and assayed for BCAT1/ IKZF1 methylation. Age-adjusted comparisons of ctDNA positivity were made with blood results from N = 310 pre-treatment CRC patients (60% male), as well as male (N = 344) and female (N = 383) controls with a clear colonoscopy and no history of cancer. Results: BCAT1 and IKZF1 were significantly hypermethylated in PCa and CRC tumors compared to matched normal tissues (Table). Conversely, BCAT1/ IKZF1 CpG probes were hypomethylated in breast tumors compared to matched normal samples. There was no correlation between hypermethylation in IKZF1/ BCAT1 and stage of PCa or CRC. In contrast to PCa in silico findings of hypermethylation, BCAT1/IKZF1 ctDNA methylation was only positive in 7/101 PCa blood samples (6.9%; 95%CI 3.2-13.9%). Three of 32 BCa blood samples (9.4%; 95%CI 2.5-25%) were positive for BCAT1/IKZF1 methylation. These positivity rates were not significantly different to controls at 8.5% for females (p = 0.80 vs BCa) and 8.2% for males (p = 0.73 vs PCa), but all were significantly lower than that observed in CRC cases (59.0%, 183/310; p < 0.001). Conclusions: While interrogation of TCGA datasets revealed IKZF1 and BCAT1 hypermethylation in prostate tumors, detection rate of these biomarkers in blood from PCa patients was not higher than that of non-cancer controls. Likewise, the BCAT1/IKZF1 ctDNA assay positivity rate in BCa patients was also no different to non-cancer controls. These findings highlight the specificity of methylated BCAT1 and IKZF1 ctDNA for detection of CRC.[Table: see text]
Predicting biomarkers of hepatic metastasis in Chinese colorectal cancer patients.
Authors: Peng Honghua , Mu Tianhao , Sheng Yaping , Li Yingmei , Cao Peiguo 
Published: June 2021

e15545 Background: Hepatic metastasis is the most common site of distant spread from colorectal cancer. About 15-25% patients with colorectal cancer harbors hepatic metastasis. The molecular mechanism and predicting biomarkers in colorectal cancer are still not fully understood. Methods: 57 Chinese colorectal cancer patients were enrolled in a cohort study. Samples of primary tumor were collected in these patients and underwent whole exome sequencing. Mutation profiles of primary tumors between the patients with metastasis and those without metastasis were analyzed and compared. Results: In the cohort, 54.4% (31/57) patients presented hepatic metastasis at the time of diagnosis, while 45.6% (26/57) did not. The patients were divided into 2 groups—with hepatic metastasis and without hepatic metastasis. The mutation landscape of primary tumor indicated that the Top 3 most frequently mutated genes of both groups were the same and presented mutated TP53, APC, and KRAS. 2. Interestingly, compared with the patients without hepatic metastasis, the patients with hepatic metastasis presented a higher frequency of mutated TCF7L2 (35.5% vs 3.85%) and TRIM77 (16.1% vs 0%). Moreover, in the patients with hepatic metastasis, the patients with TRIM77 mutation in primary tumor showed a worse overall survival (p < 0.0001). Conclusions: TCF7L2 and TRIM77 may be identified as potential candidate predicting biomarkers for hepatic metastasis in colorectal patients. In addition, mutated TRIM 77 predicted a poor overall survival in hepatic metastasis from colorectal cancer.
Update results from ALTER-C-001: Efficacy and safety of anlotinib plus XELOX regimen as first-line treatment followed by maintenance monotherapy of anlotinib for patients with mCRC-A single-arm, multicenter, phase Ⅱ clinical trial.
Authors: Yan Jin , Han Yunwei , Zhang Li , Jin Yongdong , Sun Hao 
Published: June 2021

e15550 Background: The combination of anti-VEGF or anti-EGFR targeted drugs with chemotherapy is the standard first-line therapy for metastatic colorectal cancer (mCRC), and the followed maintenance treatment is an optional approach to balance the efficacy and toxicity. However, studies regarding the maintenance strategies based on antiangiogenic TKIs are limited currently. Anlotinib, a novel oral multi-target TKI which can inhibit both tumor angiogenesis and tumor cell proliferation simultaneously, substantially prolonged the PFS with manageable toxicity for refractory mCRC in the phase III ALTER0703 clinical trial. Here we report an update on the effectiveness and safety of anlotinib plus XELOX as first-line treatment followed by anlotinib monotherapy for mCRC. Methods: In this open label, single-arm, multicenter phase II clinical trial, 53 mCRC patients without prior systemic treated, aged 18-75 and an ECOG performance status of 0 or 1 were planned to recruit. Eligible patients received capecitabine (1000 mg/m2, po, d1-14, q3w) and oxaliplatin (130 mg/m2, iv, d1, q3w) plus anlotinib (10mg, po, d1̃14, q3w) treatment for 6 cycles. After 6 cycles of inducing therapy, patients would receive anlotinib (12mg, po, d1̃14, q3w) as maintenance therapy until disease progression or intolerable adverse events (AEs). The primary endpoint was PFS; Secondary endpoints included ORR, DCR, DOR and safety. Results: By the data analysis cutoff date of January 22, 2021, a total of 18 patients were enrolled, of which 12 patients were available for efficacy assessment. In best overall response assessment, there were 50.0% PR (6/12), 33.3% SD (4/12) and 16.7% PD (2/12). The ORR was 50.0% (95% CI, 21.1-78.9%) and DCR was 83.3% (95% CI, 51.5-97.9%). The longest duration of treatment was 8.8 months and the response was still ongoing. The median PFS was not reached. The most common treatment related adverse events (TRAEs) of any grade (≥20%) were leukopenia, hypertension, neutropenia, diarrhea, fatigue, hypertriglyceridemia. Grade 3/4 TRAEs included hypertension (22.2%), hypertriglyceridemia (11.1%), lipase elevated (11.1%) and neutropenia (5.6%). No grade 5 AEs occurred. Conclusions: The update results suggested that anlotinib combined with XELOX as first line regimen followed by anlotinib monotherapy showed promising anti-tumor activity and manageable safety for patients with mCRC. And the conclusions needed to be confirmed in trials continued subsequently. Clinical trial information: ChiCTR1900028417.
The temporal evaluation of RAS and BRAF mutation by liquid biopsy at progression after bevacizumab combinations in patients with metastatic colorectal cancer (mCRC).
Authors: Bilgetekin Irem , Dogan Mehmet , Karacin Cengiz , Basal Fatma Bugdayci , Esin Ece , Ucar Gokhan , Isak Ozlem Aydin , Imamoglu Goksen Inanc , Yildiran Keskin Gul Sema , Erturk Ismail , Aktas Burak Yasin , Kose Baytemur Naziyet , Aydin Kubra , Ozturk Selcuk Cemil , Aksel Bulent , Karadurmus Nuri , Kandemir Olcay , Oksuzoglu Berna , Demirci Umut 
Published: June 2021

e15587 Background: Expanded RAS analysis is essential for the selection of biologic agents in mCRC. RAS mutations indicates anti-EGFR unresponsiveness. In this study, we aimed to investigate RAS and BRAF mutations by liquid biopsy at progression in patients with RAS mutant mCRC. Methods: Sixty patients with mCRC who harbored tissue RAS mutations were prospectively analyzed between July 2019 and April 2020. All the patients treated with chemotherapy plus bevacizumab combinations . The plasma samples of the patients were analyzed after progression of bevacizumab combinations. RAS mutation profile was evaluated in plasma using Idylla PCR-based molecular diagnostics method, which enables rapid detection of common mutations in RAS and BRAF genes in circulating tumor DNA (ctDNA). Kaplan-Meier method was used for survival analysis and log-rank test was performed for comparison of groups. Results: The median age of the patients was 60 years (IQR:35-83 years) and female was (n=23, 38.3%). Primary tumor was located in the left colon in 81.7% of all patients. There were 95.0% KRAS and 5% NRAS mutations in baseline tissue biopsy. As a result of liquid biopsy after progression, 55.0% of the patients had KRAS, 3.3% NRAS and 3.3% had BRAF mutations. The RAS mutation detected in 58.3% of the patients. While there was no significant difference in terms of clinicopathological features between wild type (RAS/BRAF) and mutant type (RAS/BRAF) determined by liquid biopsy, the overall survival (OS) of the wild type group was significantly longer than mutant group (43.8 vs. 20.4 months, p= 0.002). Conclusions: This study demonstrated that there may be changes in RAS/BRAF mutation from plasma analysis after progression in patients with mCRC. Since better survival in the patient group with wild type was detected compared to the RAS concordance group, the evaluation of RAS mutation status at the time of progression may be important in terms of disease prognosis and treatment options.
Rationale for the utilization of biological targeted agents (bTAs) in the treatment of metastatic colorectal cancer (mCRC), single tertiary cancer center experience.
Authors: Barrera Ivan , Lakehal Yahia A. , Kavan Tomas , Kavan Petr 
Published: June 2021

e15589 Background: Worldwide treatment for 1st line (1LTx) mCRC included doublet or triple chemotherapy with or without bTAs. In Quebec, the anti-EGFR therapy (panitumumab or cetuximab) was only recently approved in this setting for patients RAS WT, Bevacizumab (B) not eligible. In this study we evaluated the rational and outcomes using bTAs. Methods: Retrospective study assessing mCRC pts treated with or without bTAs at any time throughout the course of therapy at the Jewish General Hospital between 2010-2018. Pts were divided in 3 groups according to their 1LTx for analysis: Chemotherapy alone (1LChA), Chemotherapy plus B (1LChB), and anti-EGFR with or without chemotherapy (1LaEGFR). The primary objective was to assess the rational of bTAs prescription based in 1LTx selection. Secondary objectives included safety, PFS and OS. Results: Among a total of 463 pts with mCRC; 196 pts (42.3%) received 1LChA, 246 pts (53.2%) 1LChB, and 21 pts (4.5%) 1LaEGFR respectively. 1LChA group, 51% omitted bTAs for physician-patient preferences, 96 pts (49%) had contraindications for B, and 79 pts (40.3%) were potentially candidates for aEGFR, but did not receive it. 152/196 (77.5%) pts continued to 2LTx and 34.8% received bTAs. As for the 3LTx, 78/196 (40%) received a treatment, 48.7% received bTAs. The most common grade 3-4 adverse events (AEs) were hypertension and bowel perforation in B, gastrointestinal (GI) symptoms and skin reaction (SR) in aEGFR. 1LChB group, 31 pts (12.6%) presented AEs related to B. 191/246 pts (77.6%) continued to 2LTx with 48 pts (25%) receiving ChB despite progression in 1LTx on this bTA and 19 pts (10%) receiving aEGFR. The most common AEs reported in 2LTx were GI symptoms and neuropathy. In 3LTx, 54/91pts (59.3%) received aEGFR therapy and 8 pts (14.8%) had SR AEs. 46 pts (18.6%) continued to 4LTx, 13/46 pts (28.2%) received aEGFR. 1LaEGFR group, the most common AEs were SR and GI symptoms. 11/21 pts (52.3%) continued to 2LTx; 5 pts (45.4%) switching bTA class and receiving ChB. 81% pts started treatment between 2017-2018 and had at least two contraindication criteria for. The median PFS for the 1LChA and 1LChB groups were 10 and 11.5 months, respectively, and were not statistically significant (p=0.22). The OS with 1LChA and 1LChB was 33.26 vs. 27.80 months (p=0.27). The PFS and OS between 1LChA and 1LaEGFR were 10 vs 11 months (p= 0.27) and 33.26 vs. 35.07 months (p=0.13). Conclusions: The outcome and tolerability of bTAs in mCRC appear similar in our institution and randomised trials. We were not able to detect any significant difference among the three groups of comparison. The 1LaEGFR available data in this subset of patients are limited. Our data highlights the importance of optimal therapeutic sequencing to prolong OS. Dedicated studies are needed in order to determine the best bTAs therapeutic strategy in mCRC.