| Cervical cancer is one of the most lethal malignancy in developing countries for women.To a certain extent,acquired resistance of cervical cells to chemotherapeutic agent is one of reasons.However,the molecular bases are not well understood.In our study,we developed cisplatin resistance in parental cell line,Hela cells,and subsequently examined the molecular mechanisms linked to the acquired resistance.Transcriptome sequencing was utilized to compare the expression patterns of the human cervical adenocarcinoma Hela cisplatin-resistant cell line Hela/DDP with those of its progenitor cell line Hela.The present study has found 2,312 DEGs,which including 1,437 up-regulated genes and 875 down-regulated ones.Gene ontology(GO),Cluster of Orthologous Groups of proteins(COG)and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway analysis were performed to reveal potential molecular mechanisms.According to KEGG database,the most genes were enriched in PI3K/AKT signal pathway in Hela/DDP cells.The validation of q-PCR showed that six upregulated DEGs were WNT10 B,INHBB,CACNA1 H,HSP90B1,CREB5,and AKT3,whereas eight downregulated DEGs were SIPA1L1,FLNC,LAMA4,CD70,PDGFC,CREB3L1,COL4A6 and IL1 A respectively.AKT3 is a key factor in the PI3K/AKT signaling pathway.And it was highly expressed in Hela/DDP cells.The si RNA interference assay and inhibitor treatment in Hela/DDP enhanced the sensitivity to cisplatin.q RT-PCR experiments showed that the expression of Akt3 was decreased after RNAi interference and inhibitor treatment.Western Blot showed that the expression of p AKT was inhibited.CCK-8 experiments showed that low expression of AKT3/p AKT enhanced the sensitivity of drug-resistant cells to cisplatin.Apoptosis results demonstrated that inhibition of AKT3 expression increased the rate of He La/DDP apoptosis.Based on the above experimental results,one of the molecular mechanisms underlying the high expression of AKT3 or resistance to cisplatin in cervical cancer cells may be found in this study.It may provide a certain experiment fundament for inhibiting or reversing the cisplatin resistance of cervical cancer. |
