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目的研究 BRMS1基因转染卵巢上皮性癌(卵巢癌)细胞后对其体内外转移的抑制作用。方法应用脂质体介导法将 BRMS1基因转染入卵巢癌细胞株 A2780细胞内(转染组),设未进行任何转染的 A2780细胞为空白对照组,转染空载体 pcDNA3质粒者为阴性对照组。体外观察转染前后3组细胞增殖、凋亡、细胞间缝隙通讯连接、黏附转移情况及超微结构的变化。建立裸鼠人卵巢癌移植瘤模型,观察3组裸鼠 BRMS1基因抑制肿瘤体内转移的作用。结果 BRMS1基因成功转染入 A2780细胞中。BRMS1基因转染后,空白对照组细胞生长速度与阴性对照组和转染组比较,差异无统计学意义(P>0.05);空白对照组、阴性对照组及转染组细胞的每孔克隆数分别为(42±7)、(39±4)及(40±4)个,3组间比较,差异无统计学意义(P>0.05);3组细胞 S 期和 G_2/M 期、G_0/G_1期比例比较,差异也无统计学意义(P>0.05)。转染组细胞间缝隙通讯连接功能增强。细胞侵袭实验显示,转染组转入底层膜的细胞数[(112±23)个]明显低于空白对照组和阴性对照组[分别为(306±49)、(322±91)个;P<0.01]。BRMS1基因转染后,裸鼠体内转移灶,转染组为(23±7)个,分别与空白对照组和阴性对照组[分别为(96±12)、(112±20)个]比较,差异均有统计学意义(P<0.01)。结论作为转移抑制基因,BRMS1基因可抑制卵巢癌细胞的转移。
Objective To study the inhibitory effect of BRMS1 gene on the metastasis of ovarian epithelial carcinoma (ovarian cancer) cells in vitro and in vivo. Methods Transfection of BRMS1 gene into ovarian cancer cell line A2780 (lipofectamine 2000) was performed by liposome-mediated method. A2780 cells without any transfected cells were blank control group, transfected with empty vector pcDNA3 plasmid was negative Control group. In vitro, the changes of cell proliferation, apoptosis, cell-cell gap junction, adhesion and metastasis and ultrastructure were observed before and after transfection. The nude mice model of human ovarian cancer xenografts was established, and the BRMS1 gene expression in nude mice was observed to inhibit the metastasis in vivo. Results The BRMS1 gene was successfully transfected into A2780 cells. After transfection of BRMS1 gene, the growth rate of blank control group was not significantly different from that of negative control group and transfection group (P> 0.05). The numbers of clones per well of blank control group, negative control group and transfected group (42 ± 7), (39 ± 4) and (40 ± 4), respectively. There were no significant differences among the three groups (P> 0.05) G_1 phase ratio, the difference was not statistically significant (P> 0.05). Transfection group cell gap communication connectivity enhanced. Cell invasion assay showed that the number of cells transfected into the basal membrane [(112 ± 23)] was significantly lower than that of the blank control group [(306 ± 49), (322 ± 91), P <0.01]. After transfection with BRMS1 gene, the number of metastatic cells in nude mice was 23 ± 7, which were (96 ± 12) and (112 ± 20), respectively, compared with the blank control group and the negative control group The differences were statistically significant (P <0.01). Conclusion As a metastasis suppressor gene, BRMS1 gene can inhibit the metastasis of ovarian cancer cells.