Li Mei, Li Z Hubin, Song Juanrong, Li Xu, Zhai Pengtao, Mu Xudong, Qiu Fakai, Yao Le
Department of Minimally Invasive Intervention, Shaanxi Provincial Cancer Hospital, Xi'an, Shaanxi, China.
Department of Oncology, Shaanxi Provincial Cancer Hospital, Xi'an, Shaanxi, China.
Cell J. 2022 Mar;24(3):112-119. doi: 10.22074/cellj.2022.7231.
The aim of the recent study was to investigate the effects of on reversing Doxorubicin (DOX) resistance, as chemotherapeutic agents through up-regulation of in human liver cancer HepG2 cells.
In this experimental study, the drug resistance in liver cancer cells via drug efflux inhibition and enhancing apoptosis by the regulation of and multi-drug resistance/ P-glycoprotein (MDR/P-gp) expression was revealed. Using 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, effect of DOX on cell proliferation was evaluated after transfection in HepG2 and HepG2/DOX cells. Activity of P-gp on drug efflux was measured by the Rhodamine 123 (Rho-123) assay. mRNA expression levels were measured by quantitative reverse transcription polymerase chain reaction (qRT-PCR) and flow cytometry was used to measure the apoptotic ratio of HepG2/DOX cells.
overexpression considerably inhibited the HepG2/DOX cells viability (P<0.05). qRT-PCR results revealed that is a pivotal regulator in PI3K/Akt/P-gp axis. Overexpression resulted in up-regulation and ultimately down-regulation of P-gp. This inhibits drug resistance, proliferation and induces apoptosis in HepG2/DOX cells (P<0.05). Whilst, treatment with 10 μM of special inhibitors, including LY294002 (PI3K) or PD098059 (MAPK), increased Rho 123-associated MFI, treatment with 10 μM of SF1670 () almost abolished the effect of overexpression (P<0.05). Finally, we found that was down-regulated in HepG2/DOX cells, and its overexpression led to enhancing apoptosis with re-sensitization of HepG2/DOX cell lines to DOX through PTEN/PI3K/ Akt/MDR1 pathway.
These findings may introduce as a predictive biomarker and a potential treatment target for liver cancer therapy during MDR.
近期研究旨在探讨[具体物质]作为化疗药物通过上调人肝癌HepG2细胞中的[相关物质]来逆转多柔比星（DOX）耐药性的作用。
在本实验研究中，通过抑制药物外排以及调节[相关物质]和多药耐药/ P-糖蛋白（MDR/P-gp）表达来增强凋亡，从而揭示肝癌细胞中的耐药性。使用3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐（MTT）法，在HepG2和HepG2/DOX细胞中转染[相关物质]后评估DOX对细胞增殖的影响。通过罗丹明123（Rho-123）法测量P-gp的药物外排活性。通过定量逆转录聚合酶链反应（qRT-PCR）测量[相关物质]的mRNA表达水平，并使用流式细胞术测量HepG2/DOX细胞的凋亡率。
[相关物质]过表达显著抑制HepG2/DOX细胞活力（P<０.０５）。qRT-PCR结果显示，[相关物质]是PI3K/Akt/P-gp轴中的关键调节因子。[相关物质]过表达导致[相关物质]上调并最终使P-gp下调。这抑制了HepG2/DOX细胞的耐药性、增殖并诱导凋亡（P<０.０５）。同时，用10μM的特殊抑制剂处理，包括LY294002（PI3K）或PD098059（MAPK），增加了Rho 123相关的平均荧光强度（MFI），而用10μM的SF1670（[相关物质]）处理几乎消除了[相关物质]过表达的作用（P<０.０５）。最后，我们发现[相关物质]在HepG2/DOX细胞中下调，其过表达通过PTEN/PI3K/ Akt/MDR1途径使HepG2/DOX细胞系对DOX重新敏感并增强凋亡。
这些发现可能将[相关物质]引入作为肝癌多药耐药治疗期间的预测生物标志物和潜在治疗靶点。
