Abstract:

Objective　To investigate the effect of lncRNA FAM83H-AS1 modified by N6 methyladenosine (m6A) on the proliferation and invasion of endometrial cancer cells regulating the miR-485-5p/PAK1 axis. Methods　The endometrial cancer tissues and paired adjacent tissues from 76 patients who underwent surgical treatment in Department of Gynecology, Zhengzhou People's Hospital from February 2022 to September 2023 were collected. The expressions of lncRNA FAM83H-AS1 in the endometrial cancer tissues, adjacent normal tissues, endometrial cancer cells (HEP-1A, JEC, Ishikawa), and human normal endometrial cells (EEC) were detected by reverse transcription polymerase chain reaction (RT-PCR), respectively. The effect of m6A on lncRNA FAM83H-AS1 expression was detected by RT-PCR. The regulatory relationship of lncRNA FAM83H-AS1/miR-485-5p/PAK1 was analyzed by bioinformatics and dual luciferase assay. The cells were transfected to regulate the expression of lncRNA FAM83H-AS1 and miR-485-5p, and the cell proliferation and invasion were detected by CCK8 and Transwell. The one-way analysis of variance combined with post-Tukey event test and independent-sample t test were used for the statistical analysis. Results　The expression of lncRNA FAM83H-AS1 in the endometrial cancer tissues was higher than that in the adjacent tissues (4.8±1.3 vs.1.0±0.2, P=0.002 7). Compared with that in EEC, lncRNA FAM83H-AS1 was up-regulated in the endometrial cancer cells (HEP-1A, JEC, Ishikawa): Ishikawa (6.4±0.8, P<0.001), HEC-1A (3.2±0.5, P=0.017), and JEC (2.7±0.3, P=0.032); lncRNA FAM83H-AS1 was regulated by m6A modification. Compared with those in the sh-NC group, the cell proliferation and invasion in the sh-lncRNA FAM83H-AS1 group decreased (P<0.05). Overexpression of lncRNA FAM83H-AS1 promoted the proliferation and invasion of endometrial cells, and miR-485-5p could reverse this promotion effect. PAK1 gene was the target gene of miR-485-5p and positively correlated with lncRNA FAM83H-AS1 (r=0.73). Conclusions　LncRNA FAM83H-AS1 modified by m6A, as crRNA, competitively absorbs miR-485-5p, inhibits its expression, and up-regulates PAK1, promoting the proliferation and invasion of endometrial cancer cells probably.

Key words: Endometrial carcinoma,  , N6-methyladenosine,  , LncRNA FAM83H-AS1,  , MiR-485-5p/PAK1,  , Invasion

摘要：

目的　探讨lncRNA FAM83H-AS1被N6-甲基腺苷（m6A）修饰后介导miR-485-5p/PAK1轴调控子宫内膜癌细胞的增殖与侵袭机制。方法　收集2022年2月至2023年9月期间于郑州人民医院妇科进行手术治疗的76例患者子宫内膜癌组织及配对的癌旁组织，逆转录聚合酶链反应（RT-PCR）分别检测子宫内膜癌组织、癌旁正常组织以及子宫内膜癌细胞（HEC-1A、JEC、Ishikawa）、人正常子宫内膜细胞（EEC）中的lncRNA FAM83H-AS1表达，检测m6A对lncRNA FAM83H-AS1表达的影响。通过生物信息学、双荧光素酶实验分析lncRNA FAM83H-AS1/miR-485-5p/PAK1的调控关系。调控细胞中lncRNA FAM83H-AS1及miR-485-5p表达，通过CCK-8和Transwell检测细胞增殖和侵袭。采用单因素方差分析联合Tukey事后检验、独立样本t检验进行统计分析。结果　lncRNA FAM83H-AS1在子宫内膜癌组织中表达高于癌旁组织（4.8±1.3比1.0±0.2，P=0.002 7）；与EEC比较，FAM83H-AS1在子宫内膜癌细胞系中呈现梯度表达：Ishikawa（6.4±0.8，P<0.001），HEC-1A（3.2±0.5，P=0.017），JEC（2.7±0.3，P=0.032），且lncRNA FAM83H-AS1受m6A修饰的调控。与sh-NC组比较，sh-lncRNA FAM83H-AS1组细胞增殖和侵袭均减少（均P<0.05）。lncRNA FAM83H-AS1过表达对子宫内膜细胞增殖和侵袭有促进作用，而miR-485-5p能够逆转这种促进作用。PAK1基因是miR-485-5p的靶基因，与lncRNA FAM83H-AS1呈正相关（r=0.73）。结论　m6A修饰的lncRNA FAM83H-AS1作为ceRNA竞争性吸附miR-485-5p抑制其表达并上调PAK1，可能促进子宫内膜癌细胞的增殖和侵袭。

关键词:

子宫内膜癌, N6-甲基腺苷, lncRNA FAM83H-AS1, miR-485-5p/PAK1, 侵袭