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目的:探讨抑制miR-21表达对结肠癌HCT116细胞增殖、周期、凋亡、侵袭和迁移等生物学行为的影响。方法:实验分为3组,以miR-21抑制剂转染HCT116细胞为转染抑制组(IN),另设阴性对照组(NC)、空白对照组(MOCK),以Realtime PCR检测转染后HCT116细胞中miR-21的表达,应用MTT法、流式细胞术、Transwell侵袭和迁移实验检测转染后HCT116细胞的增殖、周期、凋亡、侵袭、迁移;以Western blotting检测转染后HCT116细胞PTEN的表达,荧光素酶报告实验检测转染后HCT116细胞PTEN的活性。结果:miR-21抑制剂转染后,HCT116细胞中miR-21的表达较NC和MOCK组细胞明显降低。下调miR-21后,HCT116细胞的增殖能力明显降低[72 h时:(1.05±0.45)vs(1.43±0.02),(1.45±0.01);t=13.83,P=0.000 159;t=14.88,P=0.000 119],细胞凋亡率显著增加[(16.30±1.00)%vs(1.87±0.53)%,(1.86±0.12)%;t=25.01,P=0.000 015 2;t=24.985,P=0.000 015 2],细胞周期阻滞于G0/G1期,细胞的侵袭[(50±2.0)vs(115±3.0),(111±3.0)个;t=29.09,P=0.000 008 31;t=31.23,P=0.000 006 27]和迁移能力[(22±2.0)vs(52.3±2.5),(53.0±1.0)个;t=24.01,P=0.000 017 8;t=16.34,P=0.000 082 0]明显下降。miR-21抑制剂转染的HCT116细胞中PTEN的表达及其荧光素酶相对活性均显著增加。结论:miR-21可能通过抑制PTEN进而调控大肠癌细胞生物学行为,PTEN可能是miR-21的靶基因之一。
AIM: To investigate the effects of miR-21 on the biological behavior of human colon cancer HCT116 cells such as proliferation, cycle, apoptosis, invasion and migration. Methods: The experiment was divided into three groups: HCT116 cells transfected with miR-21 inhibitor were transfection inhibition group (IN), negative control group (NC) and blank control group (MOCK) The expression of miR-21 in HCT116 cells was detected by MTT assay, flow cytometry, Transwell invasion and migration assay. The proliferation, cycle, apoptosis, invasion and migration of HCT116 cells were detected by Western blotting. PTEN expression, luciferase reporter assay test HCT116 cell PTEN activity. Results: After transfection with miR-21 inhibitor, the expression of miR-21 in HCT116 cells was significantly lower than that in NC and MOCK groups. The proliferation of HCT116 cells was significantly reduced after miR-21 was down-regulated [at 72 h: (1.05 ± 0.45) vs (1.43 ± 0.02), (1.45 ± 0.01), t = 13.83, P = 0.000159, t = 14.88, P = 0.000 119], the apoptosis rate was significantly increased [(16.30 ± 1.00)% vs (1.87 ± 0.53)%, (1.86 ± 0.12)%, t = 25.01, P = 0.0001515, t = 24.985, P = 0.000 015 2], cell cycle arrest in G0 / G1 phase and cell invasion [(50 ± 2.0) vs (115 ± 3.0), (111 ± 3.0) t = 29.09, P = 0.000 008 31; t = 31.23 , P = 0.000 006 27], and migration ability [(22 ± 2.0) vs (52.3 ± 2.5), (53.0 ± 1.0) t = 24.01, P = 0.000 017 8, t = 16.34, P = 0.000 082 0] Significant decline. The expression of PTEN and the relative luciferase activity of HCT116 cells transfected with miR-21 inhibitor were significantly increased. Conclusion: miR-21 may regulate the biological behavior of colorectal cancer cells by inhibiting PTEN. PTEN may be one of the target genes of miR-21.