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目的研究KAI1基因对乳腺癌细胞体外增殖的抑制作用。方法应用脂质体法将pCMV-KAI1质粒转染人高转移性人乳腺癌细胞株MDA-MB-231中,免疫印迹方法检测蛋白表达;利用四甲基偶氮唑盐(MTT)法、平板克隆形成实验、体外黏附及侵袭力实验判断细胞增殖能力及黏附、侵袭力的变化。流式细胞术检测细胞生长周期的变化。结果获得了稳定表达KAI1基因的MDA- MB-231乳腺癌细胞克隆。MTT法显示,转染KAI1基因组的集落形成率(25.33%±2.36%)较转染前(43.17%±2.75%)明显降低(P<0.05)。体外黏附及侵袭力实验表明,转染组的细胞黏附侵袭能力低于未转染组和转染空载体组(P<0.05)。流式细胞术显示,转染KAI1后,G_0/G_1期细胞数量增高,从36.78%±0.61%升高至64.00%±7.56%;G_2/M期数量降低,由17.88%±0.76%降至7.63%±0.60%,差异有统计学意义。结论KAI1基因可以抑制乳腺癌细胞的增殖黏附和侵袭能力,并可能通过调节细胞周期来影响细胞的增殖。
Objective To study the inhibitory effect of KAI1 gene on the proliferation of breast cancer cells in vitro. METHODS: The pCMV-KAI1 plasmid was transfected into human highly metastatic human breast cancer cell line MDA-MB-231 by lipofectamine. The protein expression was detected by Western blotting. MTT assay, Clone formation experiment, in vitro adhesion and invasion test to determine cell proliferation and adhesion, invasiveness changes. Flow Cytometry to detect changes in cell cycle. Results Clones of MDA-MB-231 breast cancer cells stably expressing the KAI1 gene were obtained. MTT assay showed that the rate of colony formation (25.33% ± 2.36%) transfected with KAI1 gene was significantly lower than that before transfection (43.17% ± 2.75%) (P <0.05). In vitro adhesion and invasion experiments showed that the cell adhesion and invasion ability of the transfected group was lower than that of the untransfected group and the transfected empty vector group (P <0.05). Flow cytometry showed that after transfection with KAI1, the number of G 0 / G 1 phase cells increased from 36.78% ± 0.61% to 64.00% ± 7.56%, while the number of G 2 / M phase decreased from 17.88% ± 0.76% to 7.63 % ± 0.60%, the difference was statistically significant. Conclusion KAI1 gene can inhibit the proliferation, adhesion and invasion of breast cancer cells and may affect cell proliferation by regulating the cell cycle.