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目的:探讨下调宫颈癌细胞系SiHa中CD147蛋白表达对肿瘤细胞增殖成瘤性等生物学活性的影响。方法:构建重组质粒pSilencer 4.1-CMV neo/CD147 siRNA1、2,稳定转染至宫颈癌细胞系SiHa,采用半定量RT-PCR法、Western blot法检测干扰后细胞CD147及凋亡相关基因Bcl-2、caspase-3 mRNA及蛋白的表达;运用MTT法检测干扰后肿瘤细胞增殖变化;裸鼠成瘤实验分析肿瘤细胞CD147蛋白表达下调后的成瘤性。结果:与对照组相比,转染了pSilencer 4.1-CMV neo/CD147 SiRNA 1、2的SiHa中CD147及Bcl-2的mRNA、蛋白表达水平有显著下调,活性caspase-3表达增加,肿瘤增殖活性下降,裸鼠成瘤实验显示干扰后肿瘤细胞成瘤性及癌肿大小均低于对照组。结论:SiRNA转染能有效抑制宫颈癌细胞系SiHa中CD147的表达,且能抑制肿瘤细胞增殖能力,肿瘤的成瘤能力亦明显下降。
OBJECTIVE: To investigate the effect of down-regulating the expression of CD147 protein in cervical cancer cell line SiHa on the biological activity of tumor cells such as proliferation and tumorigenesis. Methods: The recombinant plasmid pSilencer 4.1-CMV neo / CD147 siRNA1,2 was constructed and transfected into SiHa cell line. The expression of CD147 and Bcl-2 gene were detected by semi-quantitative RT-PCR and Western blot. , And the expression of caspase-3 mRNA and protein. MTT assay was used to detect the proliferation of tumor cells after transfection. The tumorigenicity of CD147 protein in tumor cells was evaluated by tumorigenicity assay in nude mice. Results: Compared with the control group, the mRNA and protein expressions of CD147 and Bcl-2 in SiHa transfected with pSilencer 4.1-CMV neo / CD147 SiRNA 1,2 were significantly downregulated, the activity of caspase-3 was increased, and the tumor proliferation activity Decreased, tumor formation in nude mice showed tumor cell tumorigenicity and tumor size after interference were lower than the control group. CONCLUSIONS: SiRNA transfection can effectively inhibit the expression of CD147 in SiHa cells, and inhibit the proliferation of tumor cells. The ability of tumorigenesis is also significantly decreased.