Li Shen-Jie, Zhou Jie, Zhang Ling, Xiang Wei, Hu Qi, He Yong-Yue, Chen Li-Gang
Department of Neurosurgery, the Affiliated Hospital of Southwest Medical University, Luzhou, China.
J BUON. 2017 Mar-Apr;22(2):468-473.
To investigate the role of micro RNA-21 (miR- 21) in human glioma cells and its potential disease-causing mechanism.
jetPRIME was used to transfect the miR-21- mimics and its negative control into SWOZ2 human glioma cells. Real-time fluorescence quantitative PCR assay was used to measure differences in the expression of miR-21 in SWOZ2 glioma cells, SWOZ2-miR-21-mimics cells, and control cells. Cell counting kit-8 assay was used to measure the activity of SWOZ2 glioma cells and SWOZ2-miR-21- mimics cells, and Western blot was used to measure PTEN, p-Akt, and P-glycoprotein (P-gp).
The level of miR-21 in SWOZ2-miR-21-mimics cells was significantly higher than in SWOZ2 cells and the negative control group. Compared with SWOZ2 cells, the expression of PTEN protein in SWOZ2-miR-21 cells decreased significantly, and the expression of p-Akt and P-gp protein were significantly increased. Compared with SWOZ2 cells and the negative control group, the proliferation rate of SWOZ2-miR-21-mimics cells was significantly increased (p<0.05).The rate of apoptosis as determined by flow cytometry showed that the number of apoptotic SWOZ2-miR-21- mimics cells decreased significantly (p<0.05). Transwell assay found that the invasive ability of SWOZ2-miR-21-mimics cells increased significantly, suggesting that miR-21 can mediate the biological functions of SWOZ2 cells by inhibiting the expression of PTEN.
miR-21 may regulate the proliferation and apoptosis of human glioma cells by downregulating the expression of the PTEN protein, and miR-21 may represent a potential therapeutic target for the treatment of glioma.
探讨微小RNA-21（miR-21）在人胶质瘤细胞中的作用及其潜在致病机制。
采用jetPRIME将miR-21模拟物及其阴性对照转染至SWOZ2人胶质瘤细胞中。采用实时荧光定量PCR检测SWOZ2胶质瘤细胞、SWOZ2-miR-21模拟物细胞和对照细胞中miR-21表达的差异。采用细胞计数试剂盒-8检测SWOZ2胶质瘤细胞和SWOZ2-miR-21模拟物细胞的活性，采用蛋白质免疫印迹法检测PTEN、p-Akt和P-糖蛋白（P-gp）。
SWOZ2-miR-21模拟物细胞中miR-21水平显著高于SWOZ2细胞和阴性对照组。与SWOZ2细胞相比，SWOZ2-miR-21细胞中PTEN蛋白表达显著降低，p-Akt和P-gp蛋白表达显著升高。与SWOZ2细胞和阴性对照组相比，SWOZ2-miR-21模拟物细胞的增殖率显著升高（p<0.05）。流式细胞术检测凋亡率显示，SWOZ2-miR-21模拟物细胞凋亡数量显著减少（p<0.05）。Transwell实验发现，SWOZ2-miR-21模拟物细胞的侵袭能力显著增强，提示miR-21可通过抑制PTEN表达介导SWOZ2细胞的生物学功能。
miR-21可能通过下调PTEN蛋白表达来调节人胶质瘤细胞的增殖和凋亡，miR-21可能是治疗胶质瘤的潜在治疗靶点。
