| 引用本文: | 方芳,蔡军波,潘丹.miR-101下调c-met的表达提高宫颈癌干细胞对5-氟尿嘧啶的敏感性[J].中国现代应用药学,2017,34(4):509-514. |
| FANG Fang,CAI Junbo,PAN Dan.miR-101 Increased the Sensitivity of Cervical Cancer Stem Cells to 5-Fluorouracil by Down-regulating the Expression of C-met[J].Chin J Mod Appl Pharm(中国现代应用药学),2017,34(4):509-514. |
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| miR-101下调c-met的表达提高宫颈癌干细胞对5-氟尿嘧啶的敏感性 |
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方芳1, 蔡军波2, 潘丹3
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1.台州市立医院, 体检中心, 浙江 台州 318000;2.台州市立医院, 肿瘤外科, 浙江 台州 318000;3.台州市立医院, 妇科, 浙江 台州 318000
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| 摘要: |
| 目的 探讨miR-101与宫颈癌干细胞对5-氟尿嘧啶敏感性的关系并研究其机制。方法 流式细胞术分选Hela细胞系的肿瘤干细胞。荧光定量PCR检测Hela肿瘤干细胞中miR-101的表达水平。MTT法检测Hela肿瘤干细胞在5-氟尿嘧啶和miR-101处理下的细胞活力。Western blot试验检测5-氟尿嘧啶和miR-101对Hela肿瘤干细胞c-met表达水平，PI3K、AKT磷酸化水平及caspases 活化水平的影响。流式细胞术检测Hela肿瘤干细胞在5-氟尿嘧啶和miR-101处理下的凋亡率。结果 Hela干细胞中miR-101的表达水平相较Hela非干细胞细胞显著下降。Hela干细胞转染miR-101后，其对5-氟尿嘧啶的敏感性显著上升。Western blot试验显示miR-101能显著降低Hela肿瘤干细胞的c-met蛋白表达水平，表明c-met是miR-101的靶点。western blot流式细胞实验结果表明在Hela肿瘤干细胞中，miR-101通过下调c-met的表达抑制PI3K和AKT的磷酸化，从而提高Hela肿瘤干细胞对5-氟尿嘧啶依赖的凋亡信号的敏感性，促进caspase-9和caspase-3的活化。结论 miR-101通过下调c-met的表达提高宫颈癌干细胞对5-氟尿嘧啶的敏感性。 |
| 关键词: miR-101 c-met 5-氟尿嘧啶 Hela肿瘤干细胞 PI3K/AKT磷酸化 |
| DOI：10.13748/j.cnki.issn1007-7693.2017.04.007 |
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| miR-101 Increased the Sensitivity of Cervical Cancer Stem Cells to 5-Fluorouracil by Down-regulating the Expression of C-met |
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FANG Fang1, CAI Junbo2, PAN Dan3
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1.Taizhou Hospital, Medical Examination Center, Taizhou 318000, China;2.Taizhou Hospital, Department of Surgical Oncology, Taizhou 318000, China;3.Taizhou Hospital, Department of Gynaecology, Taizhou 318000, China
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| Abstract: |
| OBJECTIVE To investigate the relationship between miR-101 and sensitivity of cervical cancer stem cells to 5-fluorouracil. METHODS Flow cytometry was performed to isolate the cancer stem cells in Hela cell line. RT-qPCR analysis was performed to detect the expression of miR-101 in Hela cancer stem cells. MTT assay was performed to evaluate the viability of Hela cancer stem cells which were treated with miR-101 and 5-fluorouracil. Western blot analysis was performed to detect the expression of c-met, phosphorylation of PI3K and AKT, and activation of caspases in the Hela cancer stem cells. Flow cytometry assay was performed to measure the apoptotic rate of Hela cancer stem cells. RESULTS Expression of miR-101 in Hela cancer stem cells was significantly lower than that in the Hela non-stem cells. Anti-tumor effect of 5-fluorouracil could be significantly enhanced when the miR-101 was transfected into the Hela cancer stem cells. The results of western blot showed that the expression of c-met was down-regulated by miR-101, indicating that the c-met gene was the target of miR-101. Furthermore, the results of Western blot analysis showed that miR-101 inhibited the phosphorylation of PI3K and AKT by targeting c-met. As the results, the sensitivity of Hela cancer stem cells to the 5-fluorouracil-induced apoptosis was significantly enhanced, and the activation of caspase-9 and caspase-3 was promoted. CONCLUSION miR-101 increased the sensitivity of cervical cancer stem cells to 5-fluorouracil by down-regulating the expression of c-met. |
| Key words: miR-101 c-met 5-fluorouracil Hela cancer stem cells PI3K/AKT phosphorylation |
