转染miR-101类似物后卵巢癌细胞对紫杉醇敏感性的变化及其机制

Sensitivity change of ovarian cancer cells transfected with miR-101 analogues to paclitaxel

目的观察转染miR-101类似物后卵巢癌细胞对紫杉醇的敏感性变化,并探讨其机制。方法培养卵巢癌SKOV3细胞,将SKOV3细胞分为观察组、对照组、单药组,三组均经0、6.25、25、100、400、1 600 ng/m L紫杉醇处理,观察组转染miR-101类似物,对照组转染无关序列,单药组不转染任何序列,MTT法观察各组细胞增殖能力,Annexin V-FITC/PI法测算各组细胞凋亡率。采用Western blotting法检测25 ng/m L紫杉醇注射液作用的观察组、对照组、单药组细胞DNMT3A蛋白。结果 0、6.25、25、100、400、1 600 ng/m L紫杉醇注射液作用的观察组细胞OD值分别为0.632±0.014、0.554±0.012、0.503±0.015、0.428±0.011、0.346±0.006、0.301±0.007,单药组分别为0.718±0.013、0.687±0.015、0.619±0.011、0.554±0.009、0.473±0.008、0.420±0.011,对照组分别为0.713±0.016、0.671±0.011、0.626±0.010、0.549±0.008、0.467±0.010、0.412±0.008,观察组与对照组相比,P均<0.05。0、6.25、25、100、400、1 600 ng/m L紫杉醇注射液作用的观察组细胞凋亡率分别为9.63%±0.38%、14.71%±0.64%、21.27%±0.79%、27.05%±0.92%、31.82%±1.07%、35.51%±1.63%,单药组分别为3.61%±0.24%、6.18%±0.48%、9.57%±0.42%、15.64%±0.75%、18.79%±1.12%、22.05%±1.34%,对照组分别为3.74%±0.29%、6.02%±0.35%、10.63%±0.58%、15.21%±0.84%、19.37%±1.26%、23.30%±1.18%,观察组与对照组相比,P均<0.05。观察组、对照组、单药组细胞中DNMT3A蛋白相对表达量分别为0.427±0.068、0.961±0.107,0.947±0.113,三组比较,P<0.05。结论转染miR-101类似物后卵巢癌细胞对紫杉醇的敏感性增强,miR-101可能通过靶向调控DNMT3A诱导卵巢癌细胞对紫杉醇的敏感性。

Objective To observe the sensitivity change of ovarian cancer cells transfected with miR-101 analogues to paclitaxel,and to explore its mechanism. Methods Ovarian cancer cells SKOV3 were cultivated and divided into 3groups： the observation group,control group and single drug group. Three groups were treated with 0,6. 25,25,100,400 and 1 600 ng/m L paclitaxel. The observation group was transfected with miR-101 mimics,control group was transfected with unrelated sequence and single drug group was not transfected with any sequence. MTT was conducted to observe the proliferation ability of cells in each group. Annexin V/propidium iodide staining was employed to detect the apoptosis rate in each group. Western blotting was employed to detect the protein expression of DNMT3 A in these three groups which were respectively treated by 25 ng/m L paclitaxel. Results The OD values of the observation group were 0. 632 ± 0. 014,0. 554± 0. 012,0. 503 ± 0. 015,0. 428 ± 0. 011,0. 346 ± 0. 006 and 0. 301 ± 0. 007,respectively. The OD values of the single drug group were 0. 718 ± 0. 013,0. 687 ± 0. 015,0. 619 ± 0. 011,0. 554 ± 0. 009,0. 473 ± 0. 008 and 0. 420 ± 0. 011,respectively. The OD values of the control group were 0. 713 ± 0. 016,0. 671 ± 0. 011,0. 626 ± 0. 010,0. 549 ± 0. 008,0. 467 ± 0. 010 and 0. 412 ± 0. 008,respectively. The differences were of statistical significance between the observation group and control group（ all P〈0. 05）. The apoptosis rates of the observation group were 9. 63% ± 0. 38%,14. 71% ±0. 64%,21. 27% ± 0. 79%,27. 05% ± 0. 92%,31. 82% ± 1. 07%,35. 51% ± 1. 63%,respectively. The apoptosis rates of the single drug group were 3. 61% ± 0. 24%,6. 18% ± 0. 48%,9. 57% ± 0. 42%,15. 64% ± 0. 75%,18. 79%± 1. 12% and 22. 05% ± 1. 34%,respectively. The apoptosis rates of the control group was 3. 74% ± 0. 29%,6. 02% ±0. 35%,10. 63% ± 0. 58%,15. 21 % ± 0. 84%,19. 37% ± 1. 26% and 23. 30% ± 1. 18%,respectively. The differences were of statistical significance between the observation group and control group（ all P〈0. 05）. The expression of DNMT3 A protein in the observation group,control group and single drug group were 0. 427 ± 0. 068,0. 961 ± 0. 107 and0. 947 ± 0. 113,respectively. The differences were of statistical significance among these three groups（ P〈0. 05）. Conclusion MiR-101 analog can increase the sensitivity of ovarian cancer cells to paclitaxel through targeted-regulating DNMT3 A.