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目的探讨通过重组腺病毒转导突变型 P27~(kipl)至人胆管癌细胞系 QBC_(939)中,观察对胆管癌细胞化疗敏感性的影响并探讨可能的机制。方法重组腺病毒载体 Ad-P27mt 转染人胆管癌细胞系 QBC_(939),经逆转录-聚合酶链反应(RT-PCR)、Western blot 检测 P27~(kipl)在胆管癌细胞中的表达;流式细胞术检测细胞周期。通过噻唑蓝比色法及流式细胞仪检测5-氟尿嘧啶(5-FU)、丝裂霉素(MMC)、环磷酰胺(CTX)对转染 P27~(kipl)前后的 QBC_(939)细胞生长抑制及凋亡的影响。结果 Ad-P27mt 转染人胆管癌细胞系 QBC_(939)后,5-FU、MMC 和 CTX 对 QBC_(939)细胞生长抑制率分别由41.89%、45.59%和38.91%明显升高为56.15%、55.65%和51.69%;凋亡率也由13.76%±3.03%(5-FU)、11.76%±3.99%(MMC)和10.46%±2.10%(CTX)明显升高为41.39%±4.32%(5-FU)、35.94%±2.71%(MMC)和34.46%±2.32%(CTX),差异均有统计学意义(P<0.05)。结论突变型 P27~(kipl)能显著增强胆管癌细胞对化疗药物的敏感性,为靶向调控细胞周期联合化疗药物治疗胆管癌提供了参考依据。
Objective To investigate the effect of recombinant adenovirus on the chemosensitivity of cholangiocarcinoma cells by transducing the mutant P27 kipl into human cholangiocarcinoma cell line QBC 939 and explore the possible mechanism. Methods Recombinant adenovirus vector Ad-P27mt was transfected into human cholangiocarcinoma cell line QBC_ (939). The expression of P27 kip1 in cholangiocarcinoma cells was detected by reverse transcription polymerase chain reaction (RT-PCR) and Western blot. Flow cytometry was used to detect cell cycle. The cells of QBC 939 cells transfected with P27 kipl were treated with 5-fluorouracil (5-FU), mitomycin C (MMC) and cyclophosphamide (CTX) by MTT assay and flow cytometry Growth inhibition and apoptosis. Results The inhibitory rates of 5-FU, MMC and CTX on the growth of QBC 939 cells were significantly increased from 41.89%, 45.59% and 38.91% to 56.15% after Ad-P27mt was transfected into human cholangiocarcinoma cell line QBC_ (939) 55.65% and 51.69% respectively. The apoptotic rate was also significantly increased from 13.76% ± 3.03% (5-FU), 11.76% ± 3.99% (MMC) and 10.46% ± 2.10% (CTX) to 41.39% ± 4.32% -FU), 35.94% ± 2.71% (MMC) and 34.46% ± 2.32% (CTX) respectively. The difference was statistically significant (P <0.05). Conclusions Mutant P27 kipl can significantly enhance the sensitivity of cholangiocarcinoma cells to chemotherapeutic drugs and provide a reference for the regulation of cell cycle and chemotherapy drugs in cholangiocarcinoma.