COL1A1基因干扰对人结肠癌HCT-8细胞增殖的影响

Effect of COL1A1 gene interference on proliferation of human colon cancer HCT-8 cells

目的探讨COL1A1基因干扰对人结肠癌HCT-8细胞增殖的影响。方法将COL1A1基因干扰质粒pGPU6/GFP/Neo-COL1A1(RNAi COL1A1)和阴性对照质粒pGPU6/GFP/Neo-shNC(PGNsN)瞬时转染入HCT-8细胞,并设空白对照组。转染后48～72 h,采用半定量RT-PCR法检测COL1A1基因在细胞中的转录水平;MTT法检测细胞的增殖活力;并将各组细胞接种至BALB/c小鼠上皮组织中,每隔6 d分别测量小鼠体内肿瘤大小。结果 RNAi COL1A1组HCT-8细胞COL1A1基因的mRNA转录水平较PGNsN组和空白对照组显著降低(P<0.01)。RNAi COL1A1组与PGNsN组和空白对照组比较,细胞的增殖活力有所降低,第4～7天差异有统计学意义(P<0.05),其中在第5和第6天差异极显著(P<0.01)。RNAi COL1A1组小鼠体内肿瘤大小与PGNsN组和空白对照组比较,前18 d差异无统计学意义(P>0.05);在第24天,显著小于两个对照组(P<0.05);在第30天,差异极显著(P<0.01)。结论 COL1A1基因被干扰后,能够抑制HCT-8细胞增殖,为COL1A1成为癌症治疗的候选基因提供了实验依据。

Objective To investigate the effect of COL1A1 gene interference on proliferation of human colon cancer HCT-8 cells. Methods HCT-8 cells were transiently transfected with COL1A1 RNA interfering plasmid pGPU6/GFP/Neo-COL1A l （ RNAi COL1A i ） and negative control plasmid pGPU6 / GFP / Neo-shNC （PGNsN） respectively, using those untransfected as blank control. The transcription levels of COL1A 1 mRNA in HCT-8 cells 48 ~ 72 h after transfection were determined by semi-quantitative RT-PCR, while the proliferation activity of HCT-8 cells by MTF method. The HCT-8 cells in various groups were inoculated into the epithelial tissue of BALB/c mice, and the tumor sizes were measured every 6 d. Results The transcription level of COL1A 1 mRNA in HCT-8 cells of RNAi COLIA1 group was significantly lower than PGNsN and blank control groups （P 〈 0. Ol ）. The proliferation activity of cells in RNAi COL1A I was lower than those in PGNsN and blank control group, which showed significant difference on days 4 ~ 7 （P 〈 0. 05 on days 4 - 7 and P 〈 0. 01 on days 5 and 6）. Compared with those in PGNsN and blank control groups, the tumor sizes of mice in RNAi COL1A 1 group showed no significant difference within 18 d （P 〉 0.05）, while were significantly smatl more than 24 d after inoculation （P 〈 0. 05 on day 24 and P 〈 0. 01 on day 30）. Conclusion COL1A1 ＇after interference inhibited the proliferation of HCT-8 ceils, which provided an experimental basis for serving COL1A 1 as a candidate gene for therapy of cancer.