The impact of zona pellucida protein 3 on proliferation and migration in renal clear cell carcinoma
Objective To explore the bioinformatics implications of zona pellucida protein 3(ZP3)in clear cell renal cell carcinoma(ccRCC),as well as to examine the effects of its expression on the prolif-eration and migration of ccRCC.Methods The UCSC database was utilized to analyze the differential ex-pression of ZP3 in renal clear cell carcinoma.ZP3 was overexpressed in renal clear cell carcinoma cell lines(786-O and OS-RC-2)by CRISPR/Cas9 technology,Western blotting and quantitative real-time poly-merase chain reaction(qPCR)were used to verify the overexpression.The groups were divided into control group and ZP3-overexpression group.Then,the cell counting kit-8(CCK-8)assay,plate clone formation assay,and Transwell assay were employed to observe the impact of ZP3 on the proliferation and migration in ccRCC.comparisons between groups were made by applying the one-sample t test.Results The Univer-sity of California,Santa Cruz(UCSC)database found that ZP3 was highly expressed in a variety of cancers(P＜0.05),including renal clear cell carcinoma,and the expression of ZP3 in tumor tissues was higher than normal tissues,with a statistically significant difference KIRC(Tumor:2.87±1.23,Normal:1.67±1.66,P＜0.05),ccRCC patients with high ZP3 expression had significantly poor survival[Overall surviv-al:(Logrank P＜0.05);Disease-free survival(Logrank P＜0.05)].The Western blotting results showed that overexpression of ZP3 resulted in a significantly higher absorbance than that of controls[786-O cells(31 066.13±1 078.634 vs.841.78±117.99,t=39.39,P＜0.05);OS-RC-2 cells(15 395.32±1 015.764 vs.549.35±187.78,t=20.33,P＜0.05)],and qPCR results showed that after overexpress-ing ZP3 Its absorbance was significantly higher than that of the control group[786-O cells(14.5±0.52 vs.1.0±0.09,t=35.8,P＜0.05);OS-RC-2 cells(34.3±1.7 vs.1.05±0.30,t=26.88,P＜0.05)].CCK-8 results showed that the cell proliferation capacity in ZP3-overexpressed cells was signifi-cantly higher than control cells[the absorbance of 786-O cells(3.65±0.11 vs.1.8±0.06,t=40.63,P＜0.05);the absorbance of OS-RC-2 cells(3.80±0.10 vs.1.80±0.11,t=40.38,P＜0.05)].P＜0.05)].The results of plate clone formation assay showed that cell proliferation was significantly enhanced after ZP3 overexpression[786-O cells(94±7 vs.59±2,t=6.05,P＜0.05);OS-RC-2 cells(212±9 vs.104±5,t=15.31,P＜0.05)].The results of Transwell assay showed that cell migration was signifi-cantly enhanced after ZP3 overexpression[786-O cells(504±17 vs.167±6,t=25.94,P＜0.05);OS-RC-2 cells(943±17 vs.316±28,t=26.59,P＜0.05)].The results of cell scratch assay showed that cell migration was significantly enhanced after ZP3 overexpression[786-O cells(0.56±0.07 vs.0.39±0.01,t=2.91,P＜0.05);OS-RC-2 cells(0.42±0.02 vs.0.29±0.008,t=9.63,P＜0.05)].Conclusion ZP3 expression is up-regulated in ccRCC,and ZP3 promotes the proliferation and migration of ccRCC.
Zona pellucida protein 3Renal clear cell carcinoma
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