SLFNL1-AS1/miR-218/ALKBH5 信号轴介导的m6A去甲基化修饰通过增强糖酵解促进宫颈癌顺铂耐药及恶性表型

The role of m6A RNA methylation in tumor progression is becoming increasingly prominent, but its function in cervical cancer has not been reported yet. It was found that ALKBH5, a m6A demethylase, was highly expressed in cervical cancer and was closely related to cisplatin resistance and prognosis. ALKBH5 could promote the proliferation, invasion and drug resistance of cervical cancer cells, and enhance aerobic glycolysis. Bioinformatics predict that ALKBH5 is modulated by SLFNL1-AS1/miR-218 axis. Thereafter, we hypothesize that the m6A demethylation mediated by SLFNL1-AS1/miR-218/ALKBH5 axis promotes cisplatin resistance and progression of cervical cancer by enhancing glycolysis. Based on bioinformatics, m6A Seq and seahorse energy metabolism tests, the role of ALKBH5 in cervical cancer development, the downstream mechanism of ALKBH5 in regulating glycolysis and the mechanism of upstream of SLFNL1-AS1/miR-218/ALKBH5 axis are studied from clinical samples, cells and animals in order to provide new targets and strategies for cervical cancer treatment, especially for reversing cisplatin resistance.

m6A RNA甲基化在肿瘤进展中的作用日益凸显，但是其在宫颈癌中的功能目前尚未见报道。申请人前期发现m6A去甲基化酶ALKBH5在宫颈癌中高表达且和顺铂耐药、生存预后密切相关；ALKBH5能够促进宫颈癌细胞增殖、侵袭及耐药，并能增强有氧糖酵解；生信预测ALKBH5受到lncRNA SLFNL1-AS1/miR-218信号轴的调控。据此提出科学假设：SLFNL1-AS1/miR-218/ALKBH5信号轴介导的m6A去甲基化修饰通过增强糖酵解促进宫颈癌顺铂耐药及恶性表型。本课题拟基于生物信息学、m6A Seq和seahorse能量代谢检测，从临床样本、细胞、动物多层次研究ALKBH5在宫颈癌进展中的作用；阐明ALKBH5通过m6A修饰调控糖酵解的下游作用机制；明确上游SLFNL1-AS1/miR-218/ALKBH5信号轴的调控机制，以期为宫颈癌靶向治疗尤其是逆转顺铂耐药提供新靶点和新策略

在该项目的资助下获得了以下主要研究结论：（1）ALKBH5 在宫颈癌耐药细胞系中表达上调，体内和体外实验证实异常高表达 ALKBH5 降低宫颈癌对顺铂的敏感性，促进宫颈癌顺铂耐药。（2）生物信息学和体外实验证实异常高表达 ALKBH5可以通过促进葡萄糖摄取以及乳酸生成显著增多而介导宫颈癌增殖及耐药。（3）通过生物信息学分析、FISH 实验、qRT-PCR\双荧光素酶报告基因检测和 Western blot 等实验证实了在宫颈癌中可能存在 lnc-SLFNL1-AS1/ miR-218/ALKBH5 ceRNA 调控网络的可能性。通过该项目的顺利开展，明确了在宫颈癌中通过 lnc-SLFNL1-AS1/ miR-218/ALKBH5 ceRNA 调控网络介导宫颈癌增殖和顺铂耐药的作用机制，为晚期和复发宫颈癌患者的 治疗提供了新的思路。（4）同时利用结余资金进一步探究了宫颈鳞状细胞癌和宫颈腺癌之间的基因组特征，并证实了 Alpelisib 可以通过抑制 ATR PI3K/AKT 通路，在 PIK3CA 突变的宫颈癌症细胞中显示出效果并增强了顺铂的疗效，这为宫颈癌的精准治疗提供了见解。