基于miR21的宮頸癌細(xì)胞順鉑敏感性機制研究

　　[摘要] 目的 探討miR21影響宮頸癌細(xì)胞株Hela及順鉑耐藥細(xì)胞株Hela/DDP順鉑敏感性的分子機制。 方法 利用riboFECTTM CP轉(zhuǎn)染試劑分別將成熟miR21 mimic及其陰性對照試劑NC轉(zhuǎn)染至Hela細(xì)胞，miR21 inhibitor及其陰性對照試劑NC轉(zhuǎn)染至Hela/DDP細(xì)胞，并將細(xì)胞分為mimic組、inhibitor組、相應(yīng)NC組及Blank組。Real-time PCR檢測PTEN mRNA的表達水平；流式細(xì)胞儀檢測細(xì)胞周期（PI法）及經(jīng)順鉑處理后的凋亡率（AnnexinⅤ/PI法）。 結(jié)果 Real-time PCR法檢測PTEN mRNA在Hela/DDP中低表達，為Hela的（0.410±0.046）倍（P < 0.01）；轉(zhuǎn)染mimic后，Hela中PTEN mRNA表達明顯低于NC組及Blank組（P < 0.01），NC組與Blank組比較差異無統(tǒng)計學(xué)意義（P > 0.05）；轉(zhuǎn)染inhibitor后，Hela/DDP中miR21表達明顯高于NC組及Blank組（P < 0.01），NC組與Blank組比較差異無統(tǒng)計學(xué)意義（P > 0.05）。AnnexinⅤ/PI檢測結(jié)果顯示，mimic組凋亡率與NC組、Blank組比較明顯減少（P < 0.05），inhibitor組凋亡率與NC組、Blank組比較顯著提高（P < 0.05）。PI法檢測結(jié)果顯示，mimic組S期所占比例與NC組、Blank組比較明顯增加（P < 0.05），NC組與Blank組比較差異無統(tǒng)計學(xué)意義（P > 0.05）；inhibitor組S期所占比例與NC組、Blank組比較明顯減少（P < 0.05），NC組與Blank組比較差異無統(tǒng)計學(xué)意義（P > 0.05）。 結(jié)論 PTEN mRNA在Hela/DDP中低表達，在Hela中高表達。上調(diào)miR21在Hela中的表達能明顯降低PTEN mRNA的表達，減少凋亡，增加細(xì)胞周期中S期所占比例，導(dǎo)致細(xì)胞耐藥；下調(diào)miR21在Hela/DDP中的表達能明顯增加PTEN mRNA的表達，增加凋亡率，降低細(xì)胞周期中S期所占比例，從而達到增加化療敏感性的效果。
　　[關(guān)鍵詞] 宮頸癌細(xì)胞；宮頸癌順鉑耐藥細(xì)胞；miR21；PTEN；順鉑；化療敏感性
　　[中圖分類號] R737.33 [文獻標(biāo)識碼] A [文章編號] 1673-7210（2018）02（c）-0013-06
　　Study on the mechanism of Cisplatin chemosensitivity in cervical cancer cells based on miR21
　　YUE Xiaoxue MIAO Jinwei LU Pan
　　Department of Gynecological Oncology， Beijing Obstetrics and Gynecology Hospital， Capital Medical University， Beijing 100006， China
　　[Abstract] Objective To detect the mechanism of miR21 gene on the sensitivity of Hela and Cisplatin-resistant Hela/DDP cells to Cisplatin. Methods Mature miR21 mimic and negative control （NC） miRNA were transfected into Hela cells， while miR21 inhibitor and negative control （NC） miRNA were transfected into Hela/DDP cells by riboFECTTM CP. Therefore， Hela cells were divided into mimic group， inhibitor group， NC group and blank group. Real-time PCR was used to measure the expression of PTEN mRNA in each group. The cell cycle was measured through PI method and apoptosis rate of cells after Cisplatin treatment was detected through AnnexinⅤ/PI by fluorescene activated cell sorter. Results Real-time PCR results showed that the expression of PTEN mRNA was an average of （0.410±0.046） fold higher in Hela/DDP than in Hela （P < 0.01）. The expression of PTEN mRNA in mimic group was obviously lower than those in NC group and blank group （P < 0.01）. The expression of PTEN mRNA in inhibitor group was significantly higher than those in NC group and blank group （P < 0.01）. There was no statistical difference between NC group and blank group （P > 0.05） in Hela and Hela/DDP cells. The results of Annexin Ⅴ/PI showed that the apoptosis rate of mimic group was lower than NC group and blank group （P < 0.05）， while the inhibitor group showed that the apoptosis rate was more than NC group and blank group （P < 0.05）. The of cell proportion of S period in mimic group was higher than those of NC group and blank group （P < 0.05）， and there was no significant difference between NC group and blank group （P > 0.05）. In the meantime， the cell proportion of S period in inhibitor group was less than those of NC group and blank group （P < 0.05）， and there was no significant difference between NC group and blank group （P > 0.05）. Conclusion PTEN mRNA is low expressed in Hela/DDP cells and highly expressed in Hela cells. The up-regulated expression of miR21 in Hela can significantly reduce the expression of PTEN mRNA， reduce apoptosis and increase the proportion of S phase in the cell cycle， thereby resulting in Cisplatin resistance. The down-regulated expression of miR21 in Hela/DDP can increase the expression of PTEN mRNA， increase the apoptosis rate and reduce the proportion of S phase in the cell cycle， so as to increase the chemosensitivity of Cisplatin.

相關(guān)熱詞搜索：癌細(xì)胞 敏感性 宮頸 機制 研究