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目的研究单羧酸转运蛋白(MCT)基因过表达对乳腺癌MDA-MB-231细胞凋亡、细胞周期、侵袭和转移能力的影响。方法将真核表达质粒pcDNA3.1/MCT及空质粒pcDNA3.1分别转染乳腺癌MDA-MB-231细胞。采用实时定量PCR(qRTPCR)和Western blot法分别检测转染后细胞内MCT基因的表达。Annexin V-FITC/PI染色和PI单染色结合流式细胞术检测MCT基因过表达对乳腺癌MDA-MB-231细胞凋亡、细胞周期的影响,Transwell实验检测MCT基因过表达对乳腺癌MDA-MB-231细胞侵袭能力的影响,划痕实验检测MCT基因过表达对乳腺癌MDA-MB-231细胞迁移能力的影响。结果实验组MCT的mRNA和蛋白水平明显高于转染空质粒的阴性对照组和未处理组;MCT过表达能促进乳腺癌MDA-MB-231细胞的凋亡,G2期细胞减少、S期细胞增多,同时能抑制癌细胞的侵袭和迁移能力。结论 MCT基因过表达能促进MDA-MB-231细胞的凋亡、调节细胞G2/M期检控点,抑制细胞的侵袭和迁移能力。
Objective To investigate the effect of monocarboxylate transporter (MCT) overexpression on the apoptosis, cell cycle, invasion and metastasis of breast cancer MDA-MB-231 cells. Methods Eukaryotic expression plasmid pcDNA3.1 / MCT and empty plasmid pcDNA3.1 were transfected into breast cancer MDA-MB-231 cells. Real-time quantitative PCR (qRTPCR) and Western blot were used to detect the expression of intracellular MCT gene. The effect of MCT gene overexpression on the apoptosis and cell cycle of breast cancer cell line MDA-MB-231 was detected by Annexin V-FITC / PI staining and PI staining. Flow cytometry was used to detect the effect of MCT gene overexpression on MDA- -MB-231 cells. Scratch assay was used to detect the effect of MCT gene overexpression on the migration of breast cancer MDA-MB-231 cells. Results The mRNA and protein levels of MCT in experimental group were significantly higher than those in negative control group and untreated group. MCT overexpression could promote the apoptosis of breast cancer MDA-MB-231 cells, reduce the number of cells in G2 phase and S phase cells Increase, at the same time can inhibit the invasion and migration of cancer cells. Conclusion Overexpression of MCT can promote the apoptosis of MDA-MB-231 cells, regulate the cell cycle G2 / M and inhibit the invasion and migration of cells.