摘要
目的探讨长基因间非编码RNA 02582(LINC02582)对宫颈癌细胞凋亡、迁移及放射敏感性的影响和分子机制。方法实时荧光定量PCR(RT-qPCR)检测宫颈癌细胞SiHa和子宫内膜基质细胞(ESC)中LINC02582表达水平。生物信息学工具、双荧光素酶实验分析验证LINC02582对miR-153的靶向作用。将si-NC、si-LINC02582、si-LINC02582+anti-miR-NC、si-LINC02582+anti-miR-153分别转染SiHa细胞,经电离辐射照射后运用流式细胞术、Transwell实验、集落形成实验、蛋白质印迹(Western blot)分别检测细胞凋亡、迁移、放射敏感性及相关蛋白表达水平。结果与ESC相比,SiHa细胞中LINC02582表达水平显著升高(P<0.05)。LINC02582靶向负调控miR-153表达。抑制LINC02582表达可促进辐射诱导的细胞凋亡率,降低细胞迁移能力,增强其放射敏感性,抑制神经钙黏素(N-cadherin)和半胱氨酸蛋白酶3(pro-caspase3)蛋白表达,促进上皮细胞钙黏蛋白(E-cadherin)、B细胞淋巴瘤相关X蛋白(Bax)和裂解的caspase 3(cleaved-caspase3)蛋白表达。抑制miR-153表达可逆转LINC02582低表达对SiHa细胞凋亡、迁移以及放射敏感性的影响。结论宫颈癌细胞中LINC02582高表达。抑制LINC02582表达可抑制宫颈癌细胞迁移,诱导细胞凋亡,增强其放射敏感性,其机制可能与调控miR-153有关。
Objective To investigate the effect of long intergenic noncoding RNA 02582(LINC02582) on apoptosis,migration and radiosensitivity cervical cancer cells,and further to explore its molecular mechanism.Methods Real-time fluorescence quantitative PCR(RT-qPCR) was used to detect the expression level of LINC02582 in cervical cancer cells SiHa and endometrial stromal cells(ESC).Bioinformatics tools and dual luciferase experimental analysed and verified the targeting effect of LINC02582 on miR-153.si-NC,si-LINC02582,si-LINC02582+anti-miR-NC,si-LINC02582+anti-miR-153 were transfected into SiHa cells,respectively.The transfected cells were exposed to X-ray radiation,and the apoptosis,migration,radiosensitivity and expression levels of related proteins were detected by flow cytometry,Transwell,colony formation assays and Western blot.Results Compared with ESC,the expression level of LINC02582 in SiHa cells was significantly increased(P<0.05).LINC02582 targets and the negatively regulates miR-153 expression.Inhibition of LINC02582 can promote radiation-induced apoptosis rate,reduce cell migration ability,enhance its radiosensitivity,inhibit the expression of N-cadherin and Pro-caspase3 proteins,and promote the expression of E-cadherin,B cell lymphoma associated X protein(Bax),and Cleared-caspase3 proteins.Inhibition of miR-153 can reverse the effects of LINC02582 low expression on apoptosis,migration and radiosensitivity of SiHa cells.Conclusion LINC02582 is highly expressed in cervical cancer cells.Inhibiting the expression of LINC02582 can inhibit cervical cancer cell migration,induce apoptosis,and enhance its radiosensitivity.The mechanism may be related to the regulation of miR-153.
作者
张伟
王欢
丰洁
ZHANG Wei;WANG Huan;FENG Jie(Department of Gynecology,Dezhou Maternal and Child Health Hospital,Shandong 253000,China;Department of Obstetrics,Dezhou Maternal and Child Health Hospital,Shandong 253000,China)
出处
《中国优生与遗传杂志》
2021年第5期600-604,共5页
Chinese Journal of Birth Health & Heredity
