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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 the proliferation of human colon cancer HCT-8 cells. Methods The COL1A1 gene interference plasmid pGPU6/GFP/Neo-COL1A1 (RNAi COL1A1) and the negative control plasmid pGPU6/GFP/Neo-shNC (PGNsN) were transiently transfected into HCT-8 cells and blank control group was set up. 48-72 h after transfection, the transcription level of COL1A1 gene in cells was detected by semi-quantitative RT-PCR; cell proliferation was detected by MTT assay; each group of cells was inoculated into epithelial tissues of BALB/c mice. The tumor size in mice was measured every 6 days. Results The transcription level of COL1A1 mRNA in HCT-8 cells was significantly lower in the RNAi COL1A1 group than in the PGNsN group and the control group (P<0.01). Compared with the PGNsN group and the blank control group, the cell proliferation activity of the RNAi COL1A1 group was decreased, and the difference between the 4th and 7th day was statistically significant (P<0.05), and the difference was significant on the 5th and 6th days (P< 0.01). There was no significant difference in tumor size between the PNIsN group and the blank control group in the RNAi COL1A1 group (P>0.05). On the 24th day, the tumor size was significantly smaller in the COL1A1 group than in the two control groups (P<0.05). At 30 days, the difference was extremely significant (P<0.01). Conclusion The interference of COL1A1 gene can inhibit the proliferation of HCT-8 cells and provide experimental basis for COL1A1 to become a candidate gene for cancer treatment.