Hua Fangfang, Xia Yonghua, Wang Huiling, Chen Ruixiang, Ren Yanfang, Yang Jun, Liang Wufeng
Department of Obstetrics and Gynecology, the First Affiliated Hospital, Xinxiang Medical University, Xinxiang 453100, China.
Email:
Zhonghua Zhong Liu Za Zhi. 2014 Jul;36(7):496-500.
To investigate the expression of metastasis-associated in colon cancer-1 (MACC-1) mediated by siRNA, and to study the effects of its downregulation on cell proliferation, cell cycle and invasion ability of cervical cancer SiHa cells.
MACC-1 siRNA and control siRNA were transfected into cervical cancer SiHa cells, and the expression of MACC-1 protein after transfection with MACC-1 siRNA was detected by Western blotting. The changes of cell proliferation, cell cycle and invasion ability of the SiHa cells were determined by CCK-8 kit, flow cytometry and Boyden chamber assay. The expressions of cell cycle- and invasion-related proteins were analyzed by Western blotting.
Compared with the untreated group (0.317 ± 0.023) and control siRNA group (0.309 ± 0.021), the expression of MACC-1 protein was downregulated in the MACC1 siRNA group (0.041 ± 0.006) (P < 0.05), and its downregulation significantly suppressed the cell proliferation, altered the cell cycle distribution and reduced the cell invasion ability of the SiHa cells (P < 0.05). Compared with the untreated group (0.217 ± 0.025 and 0.215 ± 0.024) and the control siRNA group (0.222 ± 0.023 and 0.207 ± 0.027), the expression of cyclin D1 and Cdk2 proteins were significantly decreased in the MACC1 siRNA group (0.076 ± 0.010 and 0.039 ± 0.007) (P < 0.05). Compared with the untreated group (0.099 ± 0.007) and control siRNA group (0.105 ± 0.012), the expression of p21 protein was significantly increased in the MACC1 siRNA group (0.676 ± 0.044) (P < 0.05). The downregulation of MACC-1 expression also evoked a decrease of expressions of MMP-2 and MMP-9 proteins and an increase of E-cadherin protein expression (P < 0.05).
MACC-1 downregulation-mediated inhibition of proliferation and decreased invasion ability of tumor cells may be closely associated with the alterations of expressions of cell cycle- and invasion-related proteins.
研究小干扰RNA（siRNA）介导的结肠癌转移相关蛋白1（MACC-1）的表达，探讨其表达下调对宫颈癌SiHa细胞增殖、细胞周期及侵袭能力的影响。
将MACC-1 siRNA和对照siRNA转染至宫颈癌SiHa细胞，采用蛋白质免疫印迹法检测转染MACC-1 siRNA后MACC-1蛋白的表达。采用CCK-8试剂盒、流式细胞术和Boyden小室实验检测SiHa细胞增殖、细胞周期及侵袭能力的变化。采用蛋白质免疫印迹法分析细胞周期及侵袭相关蛋白的表达。
与未处理组（0.317±0.023）和对照siRNA组（0.309±0.021）比较，MACC-1 siRNA组MACC-1蛋白表达下调（0.041±0.006）（P<0.05），其表达下调显著抑制SiHa细胞增殖，改变细胞周期分布，降低细胞侵袭能力（P<0.05）。与未处理组（0.217±0.025和0.215±0.024）及对照siRNA组（0.222±0.023和0.207±0.027）比较，MACC-1 siRNA组细胞周期蛋白D1（cyclin D1）和细胞周期蛋白依赖性激酶2（Cdk2）蛋白表达显著降低（0.076±0.010和0.039±0.007）（P<0.05）。与未处理组（0.099±0.007）和对照siRNA组（0.105±0.012）比较，MACC-1 siRNA组p21蛋白表达显著升高（0.676±0.044）（P<0.05）。MACC-1表达下调还引起基质金属蛋白酶2（MMP-2）和基质金属蛋白酶9（MMP-9）蛋白表达降低及E-钙黏蛋白表达升高（P<0.05）。
MACC-1表达下调介导的肿瘤细胞增殖抑制及侵袭能力降低可能与细胞周期及侵袭相关蛋白表达改变密切相关。
