Expression And Effect Of Prostaglandin D Synthetase In Clear Cell Renal Cell Carcinoma

Objective: To investigate the expression of prostaglandin D synthetase(PTGDS)in clear cell renal cell carcinoma(cc RCC)and the relationship between clinicopathological parameters,as well as the influence of overexpression of PTGDS on the biological behavior of cc RCC cell lines,and to preliminally explore the possible role and significance of PTGDS in cc RCC.Methods: 1.Multiple public databases were used to analyze the expression and prognostic effect of PTGDS gene in cc RCC.2.q-PCR and Western-blot assay were used to detect the expression of PTGDS m RNA and protein in fresh cc RCC tissue and para-cancer kidney tissue,and the differences were analyzed.3.Immunofluorescence method was used to locate the expression pattern of PTGDS protein in cc RCC.4.80 paraffin samples confirmed as cc RCC in the Department of Pathology from 2014 to 2018 were screened,and the protein expression of PTGDS gene in cc RCC tissues was detected by immunohistochemical method(IHC),and correlation analysis and overall survival analysis were conducted based on clinicopathological parameters.5.The CD4 and CD8 immune cells in the paraffin samples of cc RCC were detected by IHC,and the positive expression rates of the two immune cells were calculated to analyze their effects on the expression of PTGDS protein in cc RCC.6.cc RCC cell line 786-0 and normal renal cell line HK-2 were selected,and the expression of PTGDS m RNA in the two cell lines was detected by q-PCR.A 786-0 cell line transfected with PTGDS lentivirus was constructed,and the effects of PTGDS overexpression on the biological behavior of renal cancer cell lines were detected by CCK-8,scratch assay and cell cycle and apoptosis assay.Results: 1.Bioinformation data analysis showed that the m RNA and protein expression levels of PTGDS gene in cc RCC were lower than those in normal kidney tissue(P < 0.0001),and were associated with poor prognosis in cc RCC patients(P <0.001).PTGDS gene interaction proteins PTGS1 and PTGES also affect the expression of PTGDS in cc RCC and are associated with prognosis.2.PTGDS m RNA expression level(0.119±0.261)and protein expression level(0.057±0.573)in 11 fresh cc RCC tissues were lower than those in adjacent normal kidney tissues(0.936±0.254;4.101±5.578)(P < 0.05);3.Immunofluorescence detection showed that PTGDS were localized to the cytoplasm of tumor cells in renal carcinoma tissues and to the cytoplasm of tubular epithelial cells in normal renal tissues.4.The expression level of PTGDS protein in paraffin samples of 80 patients with renal carcinoma(0.117±0.003)was significantly lower than that in paracarcinoma tissues(0.214±0.089)(P < 0.0001).5.80 cases of cc RCC were divided into two groups by PTGDS protein expression level: relatively decreased PTGDS expression group(LG group)and significantly decreased expression group(SG group),and SG group had a higher WHO/ISUP grading(P < 0.05).However,there were no significant differences in tumor size,tumor site,vascular thrombus or tumor stage between the two groups(P > 0.05).The overall survival time of SG group was shorter than that of LG group,but there was no statistical difference(P > 0.05).The expression rate of CD4 positive cells in SG group was higher than that in LG group(P < 0.05),but there was no significant difference in CD8 positive cells expression rate between SG group and LG group(P > 0.05).6.In vitro experiments showed that the proliferation ability of cc RCC cell line 786-0(OE-PTGDS group)was weaker than that of negative control group(NC group)at all time periods after overexpression of PTGDS gene,with statistical difference(P <0.05).The cell mobility of OE-PTGDS group was significantly lower than that of NC group at different time periods(P < 0.05).The proportion of G1 phase cells in OE-PTGDS group was significantly lower than that in NC group,and the proportion of S phase cells was higher than that in NC group,the difference was statistically significant(P < 0.05).The apoptosis rate of OE-PTGDS group was significantly higher than that of NC group(P < 0.05).Conclusions: 1.m RNA and protein expression levels of PTGDS decreased in cc RCC tissues;2.The decreased level of PTGDS protein expression in cc RCC tissues was related to histological grading,and significantly decreased PTGDS protein level suggested higher WHO/ISUP grading;3.Overexpression of PTGDS in vitro can inhibit the proliferation and migration of cc RCC cells and promote cell apoptosis.4.PTGDS gene may play a certain anti-cancer role in cc RCC,and participate in the occurrence and development of cc RCC by influencing the proliferation,migration and apoptosis of renal cancer cells.