Rock2 Promotes The Proliferation,Invasion And Metastasis Of Renal Cell Carcinoma By Regulating IL-33

Background and purpose:Rock2 is a signaling protein that plays an important role in the development and metastasis of malignant tumors.Our previous studies have shown that Rock2 can promote the development of kidney cancer through multiple pathways.Interleukin 33(IL-33),a tissue-derived nuclear cytokine derived from the IL-1 family,is significantly expressed in renal cell carcinoma,but its mechanism of action on renal cell carcinoma remains unclear.Both Rock2 and IL-33 are highly expressed in renal cell carcinoma tissues,and we speculate that Rock2 may have some regulatory relationship with IL-33 secretion.The purpose of this study is to clarify the regulatory relationship between Rock2 and IL-33 secretion as well as the effect of Rock2 on the proliferation,invasion and metastasis of renal cell carcinoma,so as to provide new ideas for the treatment of renal cancer.Methods:The renal clear cell carcinoma 786-O cells with down-regulation of Rock2 were constructed by lentivirus transfection,and the R software DESeq2 was used to compare the differences in gene expression levels between the Rock2down-regulation group and the control group,and the up-regulated and down-regulated genes in the ROCK2 down-regulation group were screened out.The protein and supernatant of cells in Rock2 down-regulated group and control group were collected to detect the expression level of IL-33 in Rock2 down-regulated group.The regulatory relationship between Rock2 and IL-33 and the effect of IL-33 on786-O cells were further verified.The Rock2 knockdown group and the control group were set up,and the expression levels of IL-33 in the two groups were detected by QPCR,RNA-seq,and Western blot.Different amounts of IL-33 were added to the Rock2 down-regulation group and the control group,respectively.CCK-8 was used to detect the cell proliferation ability of each group.The Rock2 down-regulation group and control group were set as untreated group,IL-33 treated group,and IL-33 and IL-33 blocking antibody treated group,respectively.Annexin V staining was used to measure the apoptosis of Rock2 down-regulation group and control group.Transwell assay was used to detect the invasion ability of each group of cells.Results:1.Rock2 can promote the secretion of IL-33,and the secretion of IL-33 decreases after down-regulation of Rock2.2.CCK-8 cell experiments showed that Rock2 promoted the proliferation of cc RCC cells by up-regulating the secretion of IL-33,and the proliferation of IL-33 promoted cc RCC cells could be restored after down-regulation of Rock2.3.Annexin V apoptosis assay showed that Rock2 exerted anti-apoptotic effect on cc RCC cells by promoting IL-33 secretion.Down-regulation of Rock2 could induce apoptosis of tumor cells,and the anti-apoptosis effect of IL-33 could be restored after down-regulation of Rock2.4.Transwell cell invasion assay showed that IL-33 significantly increased the invasion ability of 786-O cells,and IL-33 blocking antibody inhibited the high invasion ability induced by IL-33.The invasion ability of 786-O cells was significantly reduced after Rock2 knockdown.IL-33 could also promote the invasion of 786-O cells with Rock2 knockdown,but the promoting effect was significantly smaller than that of the control group.Conclusion:1.Rock2 can promote the proliferation,invasion and metastasis of renal cancer cells and inhibit their apoptosis.2.The above effect of Rock2 in renal cancer cells is realized by promoting IL-33 secretion.3.Down-regulating Rock2 and blocking IL-33 can play an antitumor role.