应用RNAi技术研究乳腺癌转移抑制基因1对卵巢癌细胞侵袭转移的影响

Effect of BRMS1 on the invasion and metastasis of ovarian cancer cells by RNA interference

目的研究乳腺癌转移抑制基因1（BRMS1）对人类卵巢上皮性癌（卵巢癌）细胞侵袭、转移的影响。方法应用脂质体介导法将BRMS1-siRNA转染人卵巢癌细胞株OVCAR3细胞内（实验组），设转染不干扰任何基因的NC-siRNA的OVCAR3细胞为阴性对照组，未进行任何转染的为空白对照组。通过实时荧光定量PCR及免疫印迹法筛选并鉴定BRMS1基因有效沉默后，用Transwell侵袭、迁移实验检测BRMS1基因沉默前后3组细胞侵袭、迁移能力的变化。结果人卵巢癌细胞OVCAR3中BRMS1基因成功沉默。BRMS1基因沉默后，Transwell侵袭实验中，实验组转入底层膜的细胞数为（190±8．5）个，明显高于阴性对照组和空白对照组[分别为（144±7．8）个、（146±6．8）个；t=8．747，t=8．869；P=0．0001，Transwell迁移实验中，实验组转入底层膜的细胞数为（231±8．9）个，明显高于阴性对照组和空白对照组[分别为（177±9．7）个、（182±7．9）个；t=9．314，t=9．224；P=0．000]。结论BRMS1基因能抑制人类卵巢癌细胞的侵袭和转移，有望成为卵巢癌治疗的靶基因。

Objective To study the effect of BRMS1 on the invasion and metastasis of ovarian cancer cells. Methods BRMS1 small interfering RNA （BRMSl-siRNA） was transfected into human ovarian cancer cell-line OVCAR3 by liposome transfection method. The cells were divided into 3 groups： experimental group with BRMSI-siRNA, negative control group with siRNA that did not impact any gene, blank control group without any transfection. Changes of invasion and migration in the ceils were per-formed using transwell invasion and migration assay. Results BRMS1 gene was silenced in OVCAR3 cell line successfully detecting by quantitative real-time RT-PCR and immunoblotting.In transwell invasion assay,the numbers of cells in lower chamber passing through the membrane in transfected group were more than negative control group and blank control group （190±8.5, 144±7.8 and 146±5.8 respectively） （t=8.747, t=8.869, P=0.000）, while in transwell migration assay, the numbers of cells in lower chamber passing through the membrane in transfected group were more than negative control group and blank control group were （231±8.9, 177±9.7 and 182±7.9 respectively） （t=-9.314, t=9.224, P=0.000）, both with significant differences among the 3 groups. Conclusion BRMS1 gene could suppress the invasion and metastasis of ovarian cancer cells.