反义CDK4基因抑制人结肠癌细胞HT29生长

Antisense to CDK4 inhibits the growth of human colon cancer cells HT29

目的将反义CDK4导入人结肠癌细胞株HT29细胞,观察其对肿瘤细胞生长的影响。方法采用lipofectam ine转染方法转染HT29细胞,Northern印迹,W estern印迹,形态学和流式细胞术等方法用于转染效果的鉴定。结果转化细胞有反义CDK4的表达,而内源性CDK4 mRNA表达和蛋白合成下调,并且转化细胞的恶性行为及表型部分逆转,细胞生长受到抑制、软琼脂集落形成能力明显降低,同时揭示G1期阻滞。HT29-asCDK4细胞有较高的凋亡率。结论反义CDK4基因可抑制HT29细胞的生长和增殖、诱导其凋亡。为结直肠癌的基因治疗提供了一种新的可能的途径。

Objective Antisense CDK4 was transfected into human colorectal cancer cells HT29 to examine the effect of it on the growth and proliferation of HT29 cells. Methods Antisense CDK4 transfected the HT29 cells with lipofectamine. The transfected effects were verified by the Northern blot, Western blot, morphological method , flow cell measure, etc. Results The transformant cells showed the expression of exogenous antisense CDK4 mRNA, and downmodulation of endogenous CDK4 mRNA expression and inhibition of its protein synthesis. The transformant cells exhibited the partial reversion of the malignant phenotype and behavior, and that the cell growth and the ability of colony formation in soft agar were inhibited. The arrest of G1 phase was also revealed . the apoptosis rate is higher in HT29-asCDK4. Conclusion The antisense CDK4 gene can inhibited the cell growth and proliferation, and induce the cell apoptosis. It might provide a new way to gene therapy of the colorectal carcinoma.