摘要
目的探讨miR-10b通过mTOR/P70S6K信号通路对宫颈癌大鼠的作用机制。方法采用人宫颈癌HeLa细胞株构建宫颈癌大鼠模型,分为Gg组(宫颈癌大鼠)、Gm组(注射miR-10b-mimics)、Gi组(注射miR-10b-inhibitions)。采用HE染色观察癌组织变化,Western blotting、RT-PCR检测mTOR/P70S6K的mRNA和蛋白表达水平,Transwell检测HeLa细胞的侵袭能力。结果Gg组肿瘤组织出现丰富的血供,界限分明,切面呈鱼肉状,肿瘤细胞大小较为均等;Gm组肿瘤组织中有大量纤维血管形成,肿瘤细胞大小不等,胞浆着色较深;Gi组肿瘤组织中纤维血管较少,胞浆着色稍浅。Western blotting结果显示,mTOR/P70S6K蛋白表达水平为Gm组<Gg组<Gi组(P<0.05)。RT-PCR结果显示,mTOR/P70S6K mRNA水平为Gi组>Gg组>Gm组(P<0.05),miR-10b mRNA水平为Gi组<Gg组<Gm组(P<0.05)。Transwell结果显示,肿瘤细胞侵袭能力为Gi组>Gg组>Gm组(P<0.05)。结论miR-10b过表达可激活mTOR蛋白,增加P70S6K活性,抑制宫颈癌细胞的增殖和迁移,有望成为宫颈癌治疗的新靶点。
Objective To investigate the mechanism of miR-10b on cervical cancer rats through mTOR/P70S6K signaling pathway.Methods The human cervical cancer HeLa cell line was purchased and used to establish rat models of cervical cancer.Models were divided into Gg group(cervical cancer rats),Gm group(injected with miR-10b-mimics)and Gi group(injected with miR-10b-inhibitions).HE staining was applied to observe the changes of cancer tissue.Western blotting was used to detect the protein expression of mTOR/P70S6K,and RT-PCR was used to detect the mRNA levels of mTOR/P70S6K and miR-10b.Transwell was used to detect the invasive ability of HeLa cells.Results In Gg group,there were abundant blood supply,distinct boundaries,fish-like section and homogeneous size of tumor cells.In Gm group,there were a lot of fibroangiogenesis between the tumor nests,and darker cytoplasm in tumor cells with different sizes.In Gi group,there were fewer fibroangiogenesis and slightly lighter cytoplasmic staining in the tumor nests.Western blotting results showed that the expression of mTOR/P70S6K protein was the highest in Gi group,and the lowest in Gm group among the three groups(P<0.05).RTPCR results showed that the mRNA level of mTOR/P70S6K increased in the Gi group,but decreased in the Gm group when compared with that in Gg group(P<0.05).However,the mRNA level of miR-10b was decreased in the Gi group but increased in Gm group when compared with that in Gg group(P<0.05).Moreover,the Transwell invasion experiment showed that the invasive ability of cancer cells was the strongest in Gi group,but was the weakest in Gm group among the three groups(P<0.05).Conclusion Overexpression of miR-10b activates mTOR protein,increases P70S6K activity and inhibits the migration and proliferation of cervical cancer cells.It has the potential to become a new target for the treatment of cervical cancer.
作者
李婵玉
邓洁
罗剑波
黄超林
邹恋
LI Chanyu;DENG Jie;LUO Jianbo;HUANG Chaolin;ZOU Lian(The First Affiliated Hospital of Chengdu Medical College,Chengdu,Sichuan,610500,China)
出处
《肿瘤药学》
CAS
2020年第2期185-190,共6页
Anti-Tumor Pharmacy
