Abstract:

Objective To construct targeting epidermal growth factor receptor (EGFR) short hairpin RNA (shRNA) plasmid vector and transfect it into human endometrial carcinoma Ishikawa cells, evaluate the influence of RNA interference (RNAi) to the EGFR gene expression and epithelial-mesenchymal transition (EMT) in endometrial carcinoma cell. Methods shRNA plasmid targeting EGFR (pRNATEGFR shRNA) was constructed in vitro, and was transfected into Ishikawa cells with lipofectamine technique. Negative control group (without transfection), pRNAT-EGFRneg group (blank plasmid vector), pRNAT-EGFR1 group, pRNAT-EGFR2 group and pRNAT-EGFR3 group were established. RT-PCR and Western blotting were employed to detect the expression of EGFR mRNA and protein, and the expression of EMT related markers such as E-cadherin, α-catenin, N-cadherin and Vimentin in Ishikawa cells was determined before and after transfection. Results The expression of EGFR mRNA and protein in Ishikawa cells in pRNAT-EGFR2 group and pRNAT-EGFR3 group was significantly lower than that in negative control group (P<0.05), while there was no significant difference in the expression of EGFR mRNA and protein in Ishikawa cells between pRNAT-EGFR1 group and negative control group (P>0.05). After RNA interference inhibited the expression of EGFR gene in Ishikawa cells, the expression of E-cadherin and α-catenin mRNA and protein in pRNAT-EGFR2 group and pRNAT-EGFR3 group was significantly higher than that in negative control group (P<0.05), while the expression of N-cadherin and Vimentin in pRNAT-EGFR2 group and pRNAT-EGFR3 group was significantly lower than that in negative control group (P<0.01). Conclusion The inhibition of EGFR gene expression could partially reversed the expression of EMT related genes in endometrial carcinoma cells.

Key words: epidermal growth factor receptor, RNA interference, endometrial carcinoma, epithelial-mesenchymal transition