Background. Oxaliplatin is the first-choice chemotherapy method for patients with advanced colon cancer. However, its resistance leads to treatment failure for many patients. In our experiments, we aim to elucidate the associations among TRIM29 protein, mutant P53, and the resistance of colon cancer cells to oxaliplatin. Methods. HCT116 and HT-29 cells were cultured and transfected with plasmids pIRES2-ZsGreen1-TRIM29-flag. Western blot and real-time qRT-PCR were utilized to examine the protein and mRNA expressions of TRIM29 and other related markers, respectively. MTT assay was utilized to determine the cell growth rate and generate the inhibition curve. Continuous culture in low-concentration oxaliplatin was conducted to construct oxaliplatin-resistant cell lines. The coimmunoprecipitation method and immunofluorescence detection were used to examine the interaction between TRIM29 and mutant P53 protein in HT29 cells. Results. We successfully transfected pIRES2-ZsGreen1-TRIM29-flag into HCT116 and HT29 cells, which were utilized in the whole experiments. TRIM29 significantly increased the sensitivity of P53 mutant colon cancer cell HT29 to oxaliplatin. The oxaliplatin-resistant model of P53 mutant colon cancer cell HT29 was successfully constructed. TRIM29 physically bound with mutant P53 and retained it in the cytoplasm from the nucleus, which inhibited its transcription function of downstream genes such as MDR1. In addition, TRIM29 successfully reversed the resistance of HT29-OX resistant cell model to oxaliplatin. Conclusion. In mutant P53 colon cancer cell HT29, TRIM29 greatly increased the sensitivity of HT29 to oxaliplatin and reverse oxaliplatin resistance. The underlying mechanism is TRIM29 may increase the sensitivity of HT29 to oxaliplatin by blocking the transcriptional function of mutant P53, which inhibits the transcription function of its downstream gene such as MDR1.

中文翻译：

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TRIM29 逆转 P53 突变结肠癌细胞的奥沙利铂耐药性


背景。奥沙利铂是晚期结肠癌患者的首选化疗方法。然而，它的耐药性导致许多患者治疗失败。在我们的实验中，我们旨在阐明 TRIM29 蛋白、突变体 P53 与结肠癌细胞对奥沙利铂耐药之间的关联。方法。培养 HCT116 和 HT-29 细胞，并用质粒 pIRES2-ZsGreen1-TRIM29-flag 转染。利用Western blot和实时qRT-PCR分别检测TRIM29和其他相关标记物的蛋白和mRNA表达。 MTT法用于测定细胞生长速率并生成抑制曲线。在低浓度奥沙利铂中连续培养，构建奥沙利铂耐药细胞系。采用免疫共沉淀法和免疫荧光检测法检测HT29细胞中TRIM29与突变型P53蛋白的相互作用。结果。我们成功地将pIRES2-ZsGreen1-TRIM29-flag转染到HCT116和HT29细胞中，并在整个实验中使用。 TRIM29显着增加P53突变结肠癌细胞HT29对奥沙利铂的敏感性。成功构建P53突变结肠癌细胞HT29奥沙利铂耐药模型。 TRIM29与突变型P53物理结合，并将其从细胞核保留在细胞质中，从而抑制其下游基因如MDR1的转录功能。此外，TRIM29成功逆转了HT29-OX耐药细胞模型对奥沙利铂的耐药性。结论。在突变型P53结肠癌细胞HT29中，TRIM29大大增加了HT29对奥沙利铂的敏感性并逆转了奥沙利铂耐药性。 其潜在机制是TRIM29可能通过阻断突变体P53的转录功能来增加HT29对奥沙利铂的敏感性，突变体P53抑制其下游基因如MDR1的转录功能。