摘要
目的探讨长链非编码RNA DNM3OS(LncRNA DNM3OS)靶向微小RNA-134(miR-134)对肝细胞癌细胞增殖、侵袭的影响。方法采用qRT-PCR法检测不同人肝细胞癌细胞中DNM3OS和miR-134的表达水平;将HepG2细胞分为NC组、si-DNM3OS组、si-NC组、si-DNM3OS+miR-NC组和si-DNM3OS+miR-134 inhibitor组,转染培养48 h后,采用qRT-PCR法检测各转染组细胞中LncRNA DNM3OS和miR-134表达水平;采用CKK-8法和克隆形成实验测定增殖活性;Transwell实验测定细胞侵袭;流式细胞术分析凋亡;采用免疫印迹(western blot)检测上皮间质转化(EMT)相关蛋白表达;双萤光素酶报告基因实验检测LncRNA DNM3OS和miR-134的相互作用。结果与NC组比较,si-DNM3OS组和si-DNM3OS+miR-NC组细胞中DNM3OS表达水平、细胞存活率、侵袭细胞数和细胞中Vimentin、N-cadherin蛋白相对表达水平均降低(P<0.05),而细胞凋亡率和细胞中miR-134和E-cadherin蛋白表达水平升高(P<0.05);回补实验中发现,miR-134 inhibitor逆转了敲低LncRNA DNM3OS对肝细胞癌细胞增殖、侵袭的作用(P<0.05);miR-134在HepG2细胞中被LncRNA DNM3OS负靶向(P<0.05)。结论敲低LncRNA DNM3OS能够靶向调控miR-134水平升高来抑制肝细胞癌细胞的增殖及侵袭。
Objective To investigate the impacts of long non-coding r NA DNM3OS(Lncr NA DNM3OS)on the proliferation and invasion of hepatocellular carcinoma cells via targeting micror NA-134(mir -134).Methods The quantitative real-time polymerase chain reaction(qr T-PCr )was applied to detect the expression levels of lncr NA DNM3OS and mir -134 in different human hepatocellular carcinoma cells.HepG2 cells were induced with blank control,or transfected with si-DNM3OS,si-NC,si-DNM3OS+mir -NC,and si-DNM3OS+mir -134 inhibitor.After 48 hours of transfection,qr T-PCr was applied to detect the expression levels of lncr NA DNM3OS and mir -134 in cells.The proliferative activity was determined by cell counting kit-8(CCK-8)assay and clony formation assay.Cell invasion was measured by Transwell assay.Apoptosis was analyzed by flow cytometry.The protein expressions of epithelial mesenchymal transition(EMT)-related proteins were detected by Western blot.Dual luciferase reporter assay was used to detect the interaction between Lncr NA DNM3OS and mir -134.r esults Compared with those induced with blank control,HepG2 cells transfected with si-DNM3OS or si-DNM3OS+mir -NC had significantly lower expression level of DNM3OS,cell survival rate,number of invading cells,and protein expressions of Vimentin and N-cadherin(P<0.05),but significantly higher apoptotic rate and protein expressions of mir -134 and E-cadherin(P<0.05).Gain-of-function experiment showed that knockdown of mir -134 partially reversed the regulatory effect of lncr NA DNM3OS knockdown on the proliferation and invasion of hepatocellular carcinoma cells(P<0.05).Mir -134 was negatively targeted by lncr NA DNM3OS in HepG2 cells(P<0.05).Conclusion Knockdown of lncr NA DNM3OS can target and upregulate mir -134 to inhibit the proliferation and invasion of hepatocellular carcinoma cells.
作者
史成宇
范伟
王婷婷
张来香
SHI Chengyu;FAN Wei;WANG Tingting(First Department of Hepatobiliary Surgery(Department of Abdominal Oncology Surgery),Qingdao Central Hospital,University of Health and Rehabilitation Sciences,Shandong,Qingdao 266042,China;不详)
出处
《河北医药》
2025年第9期1456-1460,共5页
Hebei Medical Journal
基金
青岛市医药卫生科研指导项目(编号:2022-WJZD078)。
作者简介
通信作者:张来香,E-mail:h19cvb@163.com。
