Abstract:
Background Targeted therapy for clear cell renal cell carcinoma (ccRCC) can significantly improve the prognosis of patients with advanced ccRCC. Therefore, discovering new therapeutic targets is needed for developing new therapeutic drugs.
Objective To construct eukaryotic expression vector pcDNA3.0-Flag-CHMP4A, and detect the effects of CHMP4A on the proliferation and migration of ccRCC 786-O cells in vitro, as well as analyze the expression of CHMP4A in ccRCC and its influence on the prognosis of patients with ccRCC.
Methods The target gene CHMP4A was amplified from the breast library by PCR, and the CHMP4A was connected with the vector pcDNA3.0 by homologous recombination. 786-O cells were transfected with pcDNA3.0-Flag-CHMP4A, and then the protein expression of CHMP4A was detected by Western blot. The effects of CHMP4A on the proliferation and invasion of 786-O cells were detected by CCK-8, clone formation and scratch experiment. The ccRCC transcriptome database in TCGA was retrieved, and the expression of CHMP4A in different ccRCC stages, grades and lymphocytic metastasis groups was analyzed. The relationship between CHMP4A expression and the prognosis of ccRCC was analyzed by Kaplan-Meier method.
Results The pcDNA3.0-Flag-CHMP4A eukaryotic expression vector was successfully constructed and successfully expressed in 786-O cells, and Flag-CHMP4A inhibited the proliferation and migration of ccRCC 786-O cells in vitro (P＜0.01). TCGA database showed that the expression levels of CHMP4A mRNA were significantly correlated with stage, grade and lymph node metastasis of ccRCC patients (P＜0.01). The CHMP4A high expression group had longer disease-free survival and overall survival (P＜0.01).
Conclusion CHMP4A plays an important role in inhibiting the proliferation and migration of tumor cells, and its expression is positively correlated with the prognosis of patients with ccRCC. This research on CHMP4A provides a potential target for ccRCC therapy.
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