W Ren1, Y B Chi1, J L Sun2. 1. Department of Obstetrics and Gynecology, General Hospital of Northern Theater Command (Heping Campus), No. 5 Guangrong Street, Heping District, Shenyang, 110000, Liaoning, China. 2. Department of Obstetrics and Gynecology, General Hospital of Northern Theater Command (Heping Campus), No. 5 Guangrong Street, Heping District, Shenyang, 110000, Liaoning, China. n5243w@163.com.
Abstract
PURPOSE: To investigate the effect of shRNA-regulated S100A4 expression on the proliferation and apoptosis in KLE endometrial cancer cells. METHODS: S100A4-OVER and S100A4-shRNA were transfected into KLE endometrial cancer cells using lentiviral sh-RNA technology. Passive OVER-NC cell line and shRNA-NC cell line were used as a negative control group and non-transfected Control cell line as a blank control group. After 48 h of transfection, the expressions of S100A4 and protein were detected by real-time fluorescence quantitative PCR and Western blotting, respectively. CCK-8 detection and flow cytometer were used to detect cell proliferation and apoptosis, respectively. RESULTS: Compared with the normal control group and the negative control group, the transfection efficiency and shRNA targeting of the shRNA-interfered S100A4 gene were verified at the levels of mRNA and protein expression. The expression of the disrupted S100A4 gene at S100A4 mRNA and protein levels in endometrial cancer cells was determined. The proliferation efficiency of KLE cells in S100A4-OVER group was significantly higher than that in other four groups; the proliferation rate of S100A4-shRNA cells decreased slightly;, the apoptotic rate of KLE cells in S100A4-shRNA group increased significantly, and the apoptotic rate of KLE cells in S100A4-OVER group decreased compared with NC group. CONCLUSION: Specific regulation of S100A4 gene expression:, the enhanced expression of the S100A4 gene may promote the proliferation of KLE endometrial cancer cells; the inhibited expression of the S100A4 gene may promote the apoptosis of KLE endometrial cancer cells. S100A4 expression is closely related to the biological characteristics of endometrial cancer.
PURPOSE: To investigate the effect of shRNA-regulated S100A4 expression on the proliferation and apoptosis in KLE endometrial cancer cells. METHODS:S100A4-OVER and S100A4-shRNA were transfected into KLE endometrial cancer cells using lentiviral sh-RNA technology. Passive OVER-NC cell line and shRNA-NC cell line were used as a negative control group and non-transfected Control cell line as a blank control group. After 48 h of transfection, the expressions of S100A4 and protein were detected by real-time fluorescence quantitative PCR and Western blotting, respectively. CCK-8 detection and flow cytometer were used to detect cell proliferation and apoptosis, respectively. RESULTS: Compared with the normal control group and the negative control group, the transfection efficiency and shRNA targeting of the shRNA-interfered S100A4 gene were verified at the levels of mRNA and protein expression. The expression of the disrupted S100A4 gene at S100A4 mRNA and protein levels in endometrial cancer cells was determined. The proliferation efficiency of KLE cells in S100A4-OVER group was significantly higher than that in other four groups; the proliferation rate of S100A4-shRNA cells decreased slightly;, the apoptotic rate of KLE cells in S100A4-shRNA group increased significantly, and the apoptotic rate of KLE cells in S100A4-OVER group decreased compared with NC group. CONCLUSION: Specific regulation of S100A4 gene expression:, the enhanced expression of the S100A4 gene may promote the proliferation of KLE endometrial cancer cells; the inhibited expression of the S100A4 gene may promote the apoptosis of KLE endometrial cancer cells. S100A4 expression is closely related to the biological characteristics of endometrial cancer.
Entities:
Keywords:
Apoptosis; Endometrial cancer cells; Proliferation; S100A4; shRNA