Department of Obstetrics and Gynecology, General Hospital of Northern Theater Command (Heping Campus), No. 5 Guangrong Street, Heping District, Shenyang, 110000, Liaoning, China.
Clin Transl Oncol. 2021 Jan;23(1):148-154. doi: 10.1007/s12094-020-02406-7. Epub 2020 May 29.
To investigate the effect of shRNA-regulated S100A4 expression on the proliferation and apoptosis in KLE endometrial cancer cells.
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.
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.
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.
研究 S100A4 表达的 shRNA 调控对 KLE 子宫内膜癌细胞增殖和凋亡的影响。
采用慢病毒 shRNA 技术将 S100A4-OVER 和 S100A4-shRNA 转染至 KLE 子宫内膜癌细胞。将过表达 NC 细胞系和 shRNA-NC 细胞系作为阴性对照组，未转染的对照细胞系作为空白对照组。转染 48 h 后，采用实时荧光定量 PCR 和 Western blot 分别检测 S100A4 的表达。采用 CCK-8 检测和流式细胞术分别检测细胞增殖和凋亡。
与正常对照组和阴性对照组相比，转染效率和 shRNA 靶向 S100A4 基因干扰在 mRNA 和蛋白表达水平得到验证。确定了子宫内膜癌细胞中 S100A4 基因破坏的表达。S100A4-OVER 组 KLE 细胞的增殖效率明显高于其他四组；S100A4-shRNA 组细胞的增殖率略有下降；S100A4-shRNA 组 KLE 细胞的凋亡率明显增加，而 S100A4-OVER 组 KLE 细胞的凋亡率与 NC 组相比有所下降。
特异性调控 S100A4 基因表达：S100A4 基因表达增强可能促进 KLE 子宫内膜癌细胞的增殖；S100A4 基因表达抑制可能促进 KLE 子宫内膜癌细胞的凋亡。S100A4 表达与子宫内膜癌的生物学特征密切相关。
