摘要
目的探讨沉默间皮素(MSLN)基因对可溶性间皮素相关蛋白(SMRP)表达及对上皮性卵巢癌SKOV3细胞生长转移的影响。方法体外培养SKOV3细胞,实验分为3组,空白对照组不处理,阴性对照组转染MSLN siRNA阴性对照,沉默组转染MSLN siRNA。采用实时荧光定量PCR和Western blot检测MSLN和SMRP表达,采用MTT法和细胞集落形成实验检测细胞增殖,采用Transwell实验和细胞划痕实验检测细胞侵袭、迁移,通过腹腔注射观察肿瘤细胞在小鼠体内生长转移情况。结果与空白对照组和阴性对照组比较,沉默组MSLN mRNA(0.10±0.02)表达显著降低(P<0.05),SMRP蛋白(0.17±0.04)表达显著降低(P<0.05),第12、24、48、72小时的OD值均显著降低(P<0.05),细胞集落数目显著减少(P<0.05),穿膜细胞数目[(24.7±9.4)个]显著减少(P<0.05),细胞迁移距离[(504.3±53.6)μm]显著减小(P<0.05),肿瘤个数、瘤体最大径、肿瘤分布部位和肿瘤质量均显著减少(P<0.05)。结论沉默MSLN基因能够显著下调SMRP表达,抑制上皮性卵巢癌SKOV3细胞体内、体外的生长转移。
Objective To investigate the effect of silencing mesothelin ( MSLN) gene on soluble mesothelin - re- lated protein ( SMRP) expression and the growth and metastasis of epithelial ovarian cancer. Methods SKOV3 cells were cultured in vitro. The experiment was divided into blank control group ( ntreated),negative control group ( transfect- ed with MSLN siRNA negative control) and silent group ( transfected with MSLN siRNA). The expressions of MSLN and SMRP were detected by real - time fluorescence quantitative PCR and Western blot. Cell proliferations were detected by MTT and colony formation assay. Cell invasion and migration were detected by Transwell test and scratch test. Growth and metastasis of tumor cells in mice were observed by intraperitoneal injection. Results Compared with the blank control group and the negative control group,the expression of MSLN mRNA ( 0. 10 ± 0. 02) and SMRP protein ( 0. 17 ± 0. 04) in the silent group was significantly reduced ( P < 0. 05). The OD values in the silent group were significantly reduced at 12, 24,48 and 72 h ( P < 0. 05),and the number of cell colonies was also significantly reduced ( P < 0. 05). The number of crossed cells in the silent group ( 24. 7 ± 9. 4) was significantly reduced ( P < 0. 05). The migration distance of cells in the silent group [( 504. 3 ± 53. 6)μm]was significantly reduced ( P < 0. 05). The number of tumors,the largest diameter of tumors,the location of tumors and the weight of tumors in the silent group were significantly reduced ( P < 0. 05). Con- clusion Silencing MSLN gene can significantly down - regulate SMRP,and inhibit the growth and metastasis of SKOV3 cells in vivo and in vitro.
作者
陈春莹
刘小乐
程丽琴
付艳霞
CHEN Chun - ying;LIU Xiao - le;CHENG Li - qin;FU Yan - xia(Department of Obstetrics and Gynecology, the Eighth Affiliated Hospital,Sun Yat - sen University ( Futian,Shenzhen),Shenzhen 518033,Guangdong,China)
出处
《广东医学》
CAS
2019年第10期1376-1380,共5页
Guangdong Medical Journal
基金
深圳市科技创新委员会基金资助项目(编号:JCYJ20170818164219616)
关键词
间皮素
可溶性间皮素相关蛋白
上皮性卵巢癌
生长转移
mesothelin
soluble mesothelin - related protein
epithelial ovarian cancer
growth and metastasis
