Yang Yun, Luo Yanyan, Huang Shuting, Tao Yonghui, Li Chuanyin, Wang Chengcheng
School of Medicine, Guizhou University, Guiyang, Guizhou, China.
Department of Clinical Laboratory, Wenzhou Medical University, Wenzhou, Zhejiang, China.
Cancer Biomark. 2023;36(4):267-278. doi: 10.3233/CBM-210559.
Kidney renal clear cell carcinoma (KIRC) belongs to renal cell carcinoma which is a very aggressive malignant tumor with poor prognosis and high mortality. The MKRN family includes three members MKRN1, MKRN2 and MKRN3, which are closely related to cancers, and have been involved in many studies.
This study aimed to explore the roles of MKRN family in KIRC.
The expression of MKRNs was analyzed using the UALCAN database, prognostic analysis was performed with the GEPIA2 and Kaplan-Meier Plotter database, and correlation analysis was assessed by GEPIA2. The CCK-8 and colony formation assay were performed to detect cell proliferation, wound healing assays were performed to detect cell migration, cell cycles were detected by flow cytometry analysis, GST pull-down and co-immunoprecipitation assays were performed to detect the interaction of proteins, and the expression of MKRNs, p53 and other proteins were detect by immunoblotting analysis or quantitative PCR (qPCR).
MKRN1 and MKRN2 were lowly expressed in KIRC samples compared to the corresponding normal tissues, and KIRC patients with high levels of MKRN1 and MKRN2 showed higher overall survival (OS) and disease free survival (DFS) rates. The overexpression of MKRN1 and MKRN2 inhibited the proliferation of human KIRC cells by arresting the cell cycles, but shows little effect on cells migration. The expression of MKRN1 and MKRN2 are correlated, and MKRN1 directly interacts with MKRN2. Moreover, both MKRN1 and MKRN2 were closely correlated with the expression of TP53 in KIRC tumor, and promoted the expression of p53 both at protein and mRNA levels.
Our study suggests that MKRN1 and MKRN2 serve as tumor suppressors in KIRC, and act as promising therapeutic targets for KIRC treatment.
肾透明细胞癌（KIRC）属于肾细胞癌，是一种侵袭性很强的恶性肿瘤，预后差，死亡率高。MKRN家族包括三个成员MKRN1、MKRN2和MKRN3，它们与癌症密切相关，已参与许多研究。
本研究旨在探讨MKRN家族在KIRC中的作用。
使用UALCAN数据库分析MKRNs的表达，通过GEPIA2和Kaplan-Meier Plotter数据库进行预后分析，并通过GEPIA2进行相关性分析。进行CCK-8和集落形成试验以检测细胞增殖，进行伤口愈合试验以检测细胞迁移，通过流式细胞术分析检测细胞周期，进行GST下拉和免疫共沉淀试验以检测蛋白质的相互作用，并通过免疫印迹分析或定量PCR（qPCR）检测MKRNs、p53和其他蛋白质的表达。
与相应的正常组织相比，MKRN1和MKRN2在KIRC样本中低表达，MKRN1和MKRN2水平高的KIRC患者显示出更高的总生存率（OS）和无病生存率（DFS）。MKRN1和MKRN2的过表达通过阻滞细胞周期抑制人KIRC细胞的增殖，但对细胞迁移影响不大。MKRN1和MKRN2的表达相关，并且MKRN1直接与MKRN2相互作用。此外，MKRN1和MKRN2均与KIRC肿瘤中TP53的表达密切相关，并在蛋白质和mRNA水平上促进p53的表达。
我们的研究表明，MKRN1和MKRN2在KIRC中作为肿瘤抑制因子，是KIRC治疗有前景的治疗靶点。
