التفاصيل البيبلوغرافية
| العنوان: |
miR-7调控结直肠癌细胞对5-氟尿嘧啶敏感性的机制研究. (Chinese) |
| العنوان البديل: |
Regulating mechanism of miR-7 in sensitivity of colorectal cancer cells to 5-fluorouracil. (English) |
| المؤلفون: |
张岩, 田素礼, 周勇旭, 刘昶 |
| المصدر: |
Practical Oncology Journal; 2023, Vol. 37 Issue 4, p320-328, 9p |
| مصطلحات موضوعية: |
REVERSE transcriptase polymerase chain reaction, FLOW cytometry, PROTEINS, CELL culture, WESTERN immunoblotting, MICRORNA, APOPTOSIS, RNA, CELL physiology, MTOR inhibitors, COLORECTAL cancer, FLUOROURACIL, GENE expression, TREATMENT effectiveness, CELLULAR signal transduction, CELL survival, CELL proliferation, TRANSFERASES, CELL lines, COMBINED modality therapy, DRUG resistance in cancer cells, CASPASES |
| الملخص (بالإنجليزية): |
Objective The aim of this study was to investigate the effects of microRNA-7(miR-7)combined with 5-fluorouracil(5-FU)on proliferation and apoptosis of 5-FU resistant colorectal cancer(SW620/5-FU)cells, and to explore the regulatory mechanism of miR-7 expression in sensitivity of colorectal cancer to 5-FU. Methods Colorectal cancer tissues and adjacent tissues surgically removed from May 2019 to July 2021 were collected;Colorectal cancer HCT116, SW1116, DLD1, and SW620 cell lines, normal colon epithelial NCM460 cell line, and 5-FU resistant SW620 cell line(SW620/5-FU)were cultured as routine methods. The expression of miR-7 in cells and tissues was detected using RT-qPCR. Both SW620 cells and SW620/5-FU cells were transiently transfected with miR-7 mimic, miR-7 inhibitor, and vector, respectively. They were divided into the miR-7 vector group(vector group), miR-7 mimic group(overexpression group), miR-7 inhibitor group(inhibition group), miR-7 mimic+5-FU group combined with 5-FU, and control group. The cell proliferation and the 50% inhibitory concentration(IC50)of 5-FU were detected using the CCK-8 method. The cell apoptosis was detected by flow cytometry in each group, and Western blot was used to detect the expression of cleaved-caspase-3, Bid, mTOR, and p-mTOR proteins in each group. Results The expression of miR-7 in colorectal cancer tissues was significantly lower than that in adjacent tissues(2.1±1.28 vs. 8.4±2.49, P＜0.05). The expression of miR-7 in colorectal cancer cell lines was significantly lower than that in NCM460 cells(P＜0.05). The expression of miR-7 in SW620/5-FU cells was lower than that in SW620 cells(0.43±0.13 vs. 0.99±0.01, P＜0.05). After 48 hours of transfection, the expression of miR-7 in the mimic group was higher than that in the control group(5.14 ±1.18 vs. 0.96±0.04, P＜0.05), while the expression in the inhibitory group was lower than that in the control group(0.43±0.11 vs. 0.96±0.04, P＜0.05). Compared with the control group, the proliferation ability of SW620 cells in the overexpression group was significantly reduced, and the apoptosis rate increased(n=3, t=15.301, P=0.001), while the results in the inhibition group were opposite(n=3, t=17.610, P=0.002). The expression of cleaved-caspase-3 and Bid proteins in SW620 cells of the overexpression group were higher than those in the control group(n=3, t=14.367, P=0.008), while the expression of cleaved-caspase-3 and Bid in the inhibition group were lower than those in the control group(n=3, t=13.245, P=0.003). The IC50 values of 5-FU on SW620/5-FU cells were higher than SW620 cells(1 210 μg/mL vs. 220.6 μg/mL, P＜0.05). Compared with the control group and overexpression group, the viability of SW620 cells and SW620/5-FU cells in the miR-7 mimic+5-FU group significantly decreased by(36.33±3.85% vs. 99.30±0.75%;55.43±4.65% vs. 99.30±0.75%)(P＜0.05), and the apoptosis rate increased. The level of p-mTOR protein in SW620/5-FU cells in the miR-7 mimic+5-FU group decreased(0.23±0.04 vs. 0.82±0.05, P＜0.05). Conclusion Overexpression of miR-7 can increase the drug sensitivity of SW620/5-FU cells to 5-FU, and its mechanism may be related to the inhibition of p-mTOR. [ABSTRACT FROM AUTHOR] |
| Abstract (Chinese): |
目的 研究microRNA-7(miR-7)和5氟尿嘧啶(5-FU)联合对5-FU耐药结直肠癌(SW620/5-FU)细胞增殖、凋亡的影响,探讨miR-7表达对结直肠癌细胞5-FU敏感性的调控机制。方法 收集2019年5月—2021年7月手术切除的结直肠癌组织和癌旁组织;培养结直肠癌细胞系(HCT116、SW1116、DLD1、SW620)和正常结肠上皮细胞(NCM460),培养SW620细胞并构建5-FU耐药结直肠癌细胞(SW620/5-FU)。用RT-qPCR检测细胞和组织中miR-7的表达量。对SW620细胞和SW620/5-FU细胞均瞬时转染miR-7 mimic、miR-7 inhibitor和vector,分为miR-7 vector组(空载体组)、miR-7 mimic组(过表达组)和miR-7 inhibitor组(抑制组),以及与5-FU联合处理的miR-7 mimic+5-FU组和control组(空细胞组)。采用CCK-8法检测细胞增殖和5-FU半数抑制浓度(IC50),流式细胞术检测各组细胞凋亡情况,Western blot检测各组的cleaved caspase-3、Bid、mTOR和p-mTOR蛋白表达变化。结果 结直肠癌组织中miR-7的表达量明显低于癌旁组织(2.1±1.28 vs. 8.40±2.49,P＜0.05);结直肠癌细胞系中miR-7的表达量明显低于NCM460细胞(P＜0.05)。SW620/5-FU细胞中的miR-7的表达量低于SW620细胞(0.43±0.13 vs. 0.99±0.01,P＜0.05)。转染48 h后,miR-7 mimic组miR-7的表达量高于空载体组(5.14±1.18 vs. 0.96±0.04,P＜0.05),抑制组的表达量低于空载体组(0.43±0.11 vs. 0.96±0.04,P＜0.05)。与空载体组相比,过表达组中SW620细胞的增殖能力明显降低,细胞凋亡率增加(n=3,t=15.301,P=0.001),而抑制组结果与之相反(n=3,t=17.610,P=0.002);过表达组SW620细胞中cleaved caspase-3和Bid的表达水平均高于空载体组(n=3,t=14.367,P=0.008),而抑制组cleaved caspase-3和Bid的表达水平均低于空载体组(n=3,t=13.245,P=0.003)。5-FU对SW620/5-FU细胞的IC50高于SW620细胞(1 210 μg/mL vs. 220.6 μg/mL,P＜0.05)。与空细胞组和过表达组相比,miR-7 mimic+5-FU组的SW620细胞和SW620/5-FU细胞增殖活力均显著下降(分别为36.33±3.85% vs. 99.30±0.75%,P＜0.05;55.43±4.65% vs. 99.30±0.75%,P＜0.05),凋亡率增加;miR-7 mimic+5FU组中SW620/5-FU细胞的p-mTOR蛋白水平下降(0.23±0.04 vs. 0.82±0.05,P＜0.05)。结论 miR-7过表达可增加SW620/5-FU细胞对5-FU的药物敏感性,其机制可能与抑制p-mTOR相关。 [ABSTRACT FROM AUTHOR] |
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| قاعدة البيانات: |
Complementary Index |
