Department of Neurosurgery, The Second People's Hospital of Hunan Province, Changsha, Hunan 410007, China.
Department of General Surgery, The Second Xiangya Hospital, Central South University, Changsha, Hunan 410011, China.
Can J Gastroenterol Hepatol. 2021 Mar 22;2021:8870907. doi: 10.1155/2021/8870907. eCollection 2021.
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.
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.
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.
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.
奥沙利铂是晚期结肠癌患者的首选化疗方法。然而，其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、突变型 P53 与结肠癌细胞对奥沙利铂耐药性之间的关系。
培养 HCT116 和 HT-29 细胞，并转染质粒 pIRES2-ZsGreen1-TRIM29-flag。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。利用 MTT 测定法检测细胞生长率并生成抑制曲线。通过连续培养低浓度奥沙利铂构建奥沙利铂耐药细胞系。利用共免疫沉淀法和免疫荧光检测法检测 HT29 细胞中 TRIM29 和突变型 P53 蛋白之间的相互作用。
我们成功地将 pIRES2-ZsGreen1-TRIM29-flag 转染到 HCT116 和 HT29 细胞中，并将其用于整个实验。TRIM29 显著提高了 P53 突变型结肠癌细胞 HT29 对奥沙利铂的敏感性。成功构建了 P53 突变型结肠癌细胞 HT29 的奥沙利铂耐药模型。TRIM29 与突变型 P53 发生物理结合，并将其从细胞核内保留在细胞质中，从而抑制了其下游基因如 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。
在突变型 P53 结肠癌细胞 HT29 中，TRIM29 极大地提高了 HT29 对奥沙利铂的敏感性并逆转了奥沙利铂耐药性。其潜在机制可能是 TRIM29 通过阻断突变型 P53 的转录功能来增加 HT29 对奥沙利铂的敏感性，从而抑制其下游基因如 MDR1 的转录功能。
