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| 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 |
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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.
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