| 于 丹,伍志强,梅 倩,韩为东,丁佳佳,孟元光.干扰LRP16基因表达对人卵巢癌耐药SKOV3/DDP细胞耐药性的影响及机制研究[J].现代生物医学进展英文版,2018,(14):2606-2610. |
| 干扰LRP16基因表达对人卵巢癌耐药SKOV3/DDP细胞耐药性的影响及机制研究 |
| Effects of Downregulation of LRP16 Gene Expression on the Drug Resistance of Ovarian Cancer SKOV3/DDP Cells and Its Possible Mechanisms |
| Received:October 30, 2017 Revised:November 24, 2017 |
| DOI：10.13241/j.cnki.pmb.2018.14.002 |
| 中文关键词: LRP16 卵巢癌 耐药 SKOV3/DDP |
| 英文关键词: LRP16 Ovarian cancer Drug resistance SKOV3/DDP |
| 基金项目:国家自然科学基金项目(81071617；81001184) |
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| 中文摘要: |
| 摘要 目的：探究干扰人白血病相关蛋白16(LRP16)基因表达对人卵巢癌耐药SKOV3/DDP细胞耐药性的影响及其相关机制。方法：采用Real-time PCR和蛋白免疫印迹(WB)检测LRP16在敏感组(SKOV3细胞)、耐药组(SKOV3/DDP细胞)、LRP16干扰组(SKOV3/DDP细胞-稳定转染LRP16 shRNA质粒)和NC组(即阴性对照组，SKOV3/DDP细胞-稳定转染阴性对照质粒)细胞中的表达情况；MTT试验检测LRP16对SKOV3/DDP细胞耐药性的影响；彗星试验检测LRP16对顺铂(DDP)诱导DNA损伤的影响；流式细胞术(FCM)试验检测细胞凋亡变化；WB试验检测PTEN、p-Akt和NF-κB蛋白表达水平。结果：LRP16干扰组细胞的耐药指数(RI)值(3.19±0.21)显著低于耐药组细胞(6.84±0.37)(P<0.05)。DDP(25 μmol/L)处理24 h后，LRP16干扰组DNA损伤的细胞百分比、细胞凋亡百分比均显著低于耐药组和NC组(P均<0.05)，而耐药组和NC组比较差异无统计学意义(P>0.05)；LRP16干扰组细胞PTEN蛋白相对表达量高于耐药组和NC组(P均<0.05)，而p-Akt和NF-κB相对表达量低于耐药组和NC组(P均<0.05)。结论：干扰LRP16基因表达可逆转卵巢癌耐药SKOV3/DDP细胞的耐药性，PTEN/Akt/NF-κB可能是其中的关键信号通路。 |
| 英文摘要: |
| ABSTRACT Objective: To investigate the effects of downregulation of LRP16 gene expression on the drug resistance of ovarian cancer SKOV3/DD cells, and explore the possible mechanisms. Methods: The expressions of LRP16 in the sensitive group (SKOV3), re- sistant group (SKOV3/DDP), LRP16 interference group (SKOV3/DDP-stably transfected with the LRP16 shRNA plasmid) and NC group (SKOV3/DDP-stably transfected with the negative-control plasmid) were detected by the real-time PCR and Western blotting (WB) as- say. The influence of LRP16 expression on the drug-resistance of SKOV3/DDP was analyzed by the MTT assay. Moreover, Cisplatin (DDP)-induced DNA damage was determined by the Comet tail assay, flow cytometry (FCM) was applied to detect the cell apoptosis, and the protein levels of PTEN, p-Akt and NF-?资B were detected by the WB assay. Results: The resistance index (RI) value of LRP16 in- terference group was (3.19±0.21), which was much lower than (6.84±0.37) of the resistant group (P<0.05). At 24h post treatment of DDP (25 μmol/L), the percentage of DNA-damage cells and apoptotic rate of LRP16 interference group were significantly lower than those in the resistant group and the NC group (P<0.05), but there was no differences between the resistant group and the NC group (P>0.05). Meanwhile, the protein expression of PTEN in the LRP16 interference group was significantly lower than that in the resistant group and the NC group(P<0.05), and the protein expressions of p-Akt and NF-κB in the LRP16 interference group were significantly higher than those in the resistant group and the NC group(P<0.05). Conclusion: Downregulation of LRP16 gene expression was able to reverse the drug-resistance of SKOV3/DDP, and the PTEN/Akt/NF-κB may be the key pathway involved. |
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