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目的:分析重组人Krüppel类因子4(Krüppel-like factors 4,KLF4)基因在肝癌HepG2细胞中对化疗和光动力治疗的调节作用。方法:构建人KLF4慢病毒pWPTS-KLF4载体,应用RT-PCR、Western blotting检测pWPTS-KLF4感染后HepG2细胞中KLF4 mRNA和蛋白的表达,MTT实验检测HepG2细胞对顺铂、环磷酰胺或氟尿嘧啶耐受性的变化以及对光动力治疗敏感性的变化,罗丹明123染色检测HepG2细胞线粒体膜电位的变化。结果:成功构建慢病毒pWPTS-KLF4载体。顺铂、环磷酰胺或氟尿嘧啶处理HepG2细胞72 h后,pWPTS-KLF4感染组HepG2细胞存活率较对照组明显增高[(43.43±4.78)%vs(18.09±1.02)%;(110.51±4.58)%vs(75.23±5.92)%;(34.55±2.93)%vs(19.16±1.32)%,P<0.01]。光敏剂艾拉介导的光动力治疗后24 h,pWPTS-KLF4感染组HepG2细胞存活率较对照组明显减少[(37.16±3.26)%vs(57.24±8.01)%,P<0.01],细胞的线粒体膜电位明显降低。结论:重组人KLF4可以提高HepG2细胞化疗耐受性,增加光动力治疗敏感性。
OBJECTIVE: To analyze the regulatory effect of recombinant human Krüppel-like factors 4 (KLF4) gene in chemotherapy and photodynamic therapy in HepG2 hepatocellular carcinoma cells. Methods: The KLF4 lentivirus pWPTS-KLF4 vector was constructed and the expression of KLF4 mRNA and protein in HepG2 cells was detected by RT-PCR and Western blotting. The expression of KLF4 mRNA and protein in HepG2 cells was detected by RT-PCR and Western blotting. MTT assay was used to detect the effect of HepG2 cells on cisplatin, cyclophosphamide or fluorouracil The change of sex and the sensitivity to photodynamic therapy, the change of mitochondrial membrane potential of HepG2 cells was detected by rhodamine 123 staining. Results: The lentivirus pWPTS-KLF4 vector was successfully constructed. After treated with cisplatin, cyclophosphamide or fluorouracil for 72 h, the survival rate of HepG2 cells in pWPTS-KLF4 infected group was significantly higher than that in control group [(43.43 ± 4.78)% vs (18.09 ± 1.02)%; (110.51 ± 4.58)%, vs (75.23 ± 5.92)%; (34.55 ± 2.93)% vs (19.16 ± 1.32)%, P <0.01]. The survival rate of HepG2 cells in pWPTS-KLF4 infected group was significantly lower than that in control group (37.16 ± 3.26% vs (57.24 ± 8.01)%, P <0.01] at 24 h after IRA-mediated photodynamic therapy Mitochondrial membrane potential was significantly reduced. Conclusion: Recombinant human KLF4 can improve the chemotherapeutic tolerance of HepG2 cells and increase the sensitivity of photodynamic therapy.