目的：探讨抑制素β亚基A反义RNA1（INHBA-AS1）对宫颈癌HeLa 细胞EMT和鸟氨酸代谢途径的影响及其机制。方法：体外常规培养HeLa 细胞，实验分为10组：对照组、阴性对照（NC）组、sh-INHBA-AS1组、PluriSIn 1[硬脂酰辅酶A去饱和酶（stearyl CoA desaturase，SCD）抑制剂]组、NC+PluriSIn 1 组、sh-INHBA-AS1+PluriSIn 1 组、10058-F4（c-Myc 抑制剂）组、NC+10058-F4组、sh-INHBA-AS1+10058-F4组、sh-INHBA-AS1+OE-c-Myc 组。平板克隆实验检测各组细胞的增殖能力，FCM检测各组细胞的凋亡情况，Transwell 小室实验检测各组细胞的侵袭、迁移能力，qPCR 法检测各组细胞中INHBA-AS1、c-Myc、SCD 和EMT 相关基因（N-cadherin、TGF-β、ZEB1）mRNA 的表达，WB 法检测各组细胞中c-Myc、SCD、EMT 相关（N-cadherin、TGF-β、ZEB1）、S-腺苷-甲硫氨酸脱羧酶（SAMDC）和亚精胺/精胺N1-乙酰转移酶（SSAT）蛋白的表达，ELISA 检测各组细胞上清液中鸟氨酸脱羧酶（ODC）的含量。结果：敲减INHBA-AS1表达使HeLa 细胞的增殖、侵袭和迁移能力显著降低（均P<0.05）而细胞凋亡率显著升高（P<0.05），qPCR、WB法检测结果显示，敲减INHBA-AS1 均可显著抑制HeLa 细胞中c-Myc、SCD、N-cadherin、TGF-β、ZEB1和SAMDC的表达（均P<0.05），而促进SSAT的表达（P<0.05），并降低HeLa 细胞上清液中ODC的含量（P<0.05）。与c-Myc抑制剂和SCD抑制剂单独处理相比，其联合敲减INHBA-AS1后上述作用更加显著（均P<0.05）；与sh-INHBA-AS1组相比，进一步过表达c-Myc 后HeLa 细胞的增殖能力显著升高（P<0.05）、SCD和N-cadherin 蛋白表达水平显著升高（P<0.05）、细胞上清液中ODC含量显著升高（P<0.05）。结论：INHBA-AS1可通过c-Myc 调控SCD的表达，从而影响HeLa 细胞鸟氨酸代谢和EMT进程，进而促进HeLa细胞的增殖、侵袭和迁移能力。

Objective: To investigate the effects of inhibin subunit beta A antisense RNA 1 (INHBA-AS1) on epithelial-interstitial transformation (EMT) and ornithine metabolic pathway of cervical cancer cells and its mechanism. Methods: Cervical cancer HeLa cells were routinely cultured in vitro and the experiment was divided into 10 groups: control group, negative control (NC) group, sh-INHBA-AS1 group, PluriSIn 1 [stearyl CoA desaturase (SCD) inhibitor] group, NC+PluriSIn 1 group, sh-INHBA-AS1+PluriSIn 1 Group, 10058-F4(c-Myc inhibitor) group, NC+10058-F4 group, sh-INHBA-AS1+10058-F4 group, sh-INHBA-AS1+OE-c-Myc group. The proliferation ability of cells in each group was detected by plate cloning assay; the apoptosis of cells in each group was detected by flow cytometry; the invasion and migration abilities of cells in each group were detected by Transwell assay; the expressions of INHBA-AS1, c-Myc, SCD and EMT-related genes (N-cadherin, TGF-β and ZEB1) in cells of each group were detected by qPCR; the expressions of c-Myc, SCD, EMT-associated (N-cadherin, TGF-β and ZEB1), S-adenosine-methionine decarboxylase (SAMDC) and spermidine/spermine N1 acetyltransferase (SSAT) proteins in each group were detected by WB method, and the content of ornithine decarboxylase (ODC) in cell supernatant was detected by ELISA.Results: The proliferation, invasion and migration abilities of HeLa cells were significantly decreased after INHBA-AS1 expression was knocked down (all P<0.05). The apoptosis rate was significantly increased (P<0.05). The results of qPCR and WB assay showed that knockdown of INHBA-AS1 could significantly inhibit the expressions of c-Myc, SCD, N-cadherin, TGF-β, ZEB1, and SAMDC in HeLa cells by (all P<0.05), promote the expression of SSAT (P<0.05) and decrease the content of ODC in HeLa cell supernatant (P<0.05). These effects were more significant after further knockdown of INHBA-AS1 compared with c-Myc inhibitor and SCD inhibitor treatments (all P<0.05). Compared with sh-INHBA-AS1 group, further over-expression of c-Myc significantly increased HeLa cell proliferation (P<0.05), SCD and N-cadherin protein expression levels (P<0.05), and the content of ODCin cell supernatant (P<0.05). Conclusion: INHBA-AS1 can regulate the expression of SCD through c-Myc, participate in the ornithine metabolism and EMT process of HeLa cells, and promote the proliferation,invasion, and migration of HeLa cells.

武汉市卫健委医学科研项目（No. WX21C22）