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Archive > Volume 30 Issue 6 > 2023,30(6):497-504. DOI:10.3872/j.issn.1007-385X.2023.06.006 Prev Next
Basic Research
INHBA-AS1 affects ornithine metabolism and EMT process of cervical cancer HeLa cells through c-Myc/SCD pathway
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Abstract:
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.
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