摘要
目的探讨沉默神经鞘磷脂合成酶1(sphingomyelin synthase 1,SMS1)对卵巢癌细胞HO8910增殖、侵袭和迁移的影响。方法体外培养卵巢癌HO8910细胞株,设计siRNA-SMS1序列,转染细胞,分为阴性对照组(negative control,NC)、空白对照组(blank control,BC)、siRNA组;利用qRT-PCR技术检测各组细胞SMS1 mRNA相对表达量。MTT法检测HO8910细胞增殖情况、划痕实验检测转染后HO8910细胞迁移能力。Transwell实验检测细胞侵袭能力。薄层层析法检测HO8910细胞中SMS1活性、Cer水平。酸性神经鞘磷脂(sphingomyelin,SM)酶试剂盒、卵磷脂(phosphatidylcholine,PC)试剂盒分别检测HO8910细胞中SM、PC水平。结果卵巢癌细胞中SMS1 mRMA及蛋白相对表达量比正常卵巢细胞显著升高(P<0.05);转染48 h后,siRNA组细胞中SMS1 mRMA及蛋白相对表达量、SMS1酶活性比NC组与BC组均显著降低(P<0.05);siRNA-SMS1组A570比BC组与NC组显著降低(P<0.05);迁移指数、侵袭细胞数较BC组与NC组显著降低(P<0.05);与NC、BC组相比,siRNA组HO8910细胞中SM水平显著降低(P<0.05),Cer水平显著升高(P<0.05)。结论SMS1基因沉默可抑制卵巢癌HO8910细胞增殖、迁移及侵袭能力,可能与下调细胞中SM水平、上调Cer水平有关。
Purpose To study the effects of silencing sphingomyelin synthase-1(SMS1)on proliferation,invasion and migration of ovarian cancer cell line HO8910.Methods Ovarian cancer HO8910 cell line was cultured in vitro,and siRNA-SMS1 sequence was designed.The transfected cells were divided into negative control(NC)group,blank control(BC)group and siRNA group.The relative expression of SMS1 was detected by qRT-PCR,MTT assay was used to detect the proliferation of HO8910 cells.Scratch assay was used to detect the migration ability of HO8910 cells after transfection.Transwell assay was used to detect cell invasiveness.The activity of SMS1 and the level of Cer in HO8910 cells were detected by thin layer chromatography(TLC),and acid sphingomyelin(SM)enzyme kit and phosphatidylcholine(PC)kit were used to detect the levels of SM and PC in HO8910 cells.Results The relative expression of SMS1 mRMA and protein in ovarian cancer cells was significantly higher than those in normal ovarian cells(P<0.05).After 48 hours of transfection,the relative expression of SMS1 mRMA and protein and the activity of SMS1 enzyme in siRNA group were significantly lower than those in NC group and BC group(P<0.05).After siRNA-SMS1 transfection,the A570 of siRNA group was significantly lower than that of BC group and NC group(P<0.05).After transfection,the migration index and the number of invasive cells in siRNA group were significantly lower than those in BC group and NC group(P<0.05).After transfection,compared with NC and BC groups,SM level in HO8910 cells in siRNA group decreased significantly(P<0.05),while Cer level increased significantly(P<0.05).Conclusion SMS1 gene silencing can inhibit the proliferation,migration and invasion of ovarian cancer HO8910 cells,which may be related to down-regulation of SM level and up-regulation of Cer level.
作者
杭月
林昌岫
黄成日
HANG Yue;LIN Chang-xiu;HUANG Cheng-ri(Department of Obstetrics and Gynecology,Affiliated Hospital of Yanbian University,Yanbian 133000,China;Department of Central Laboratory,Affiliated Hospital of Yanbian University,Yanbian 133000,China)
出处
《临床与实验病理学杂志》
CAS
CSCD
北大核心
2020年第8期920-923,927,共5页
Chinese Journal of Clinical and Experimental Pathology
基金
国家自然科学基金(81360382)
吉林省教育厅“十三五”科学技术项目(JJKH20191118KJ)。
关键词
卵巢肿瘤
神经鞘磷脂合成酶1
细胞增殖
细胞迁移
ovarian neoplasms
sphingomyelin synthase 1
cell proliferation
cell migration
