Decreased Expression Of NPRL2 In Renal Cancer Cells Is Associated With Unfavourable Pathological, Proliferation And Apoptotic Features

Objective:(1)To investigate the expression of NPRL2 and the correlation with clinicopathological features in clear cell renal cell carcinoma (ccRCC);(2)To explore the function of NPRL2 in human ccRCC in cell culture models.Methods:(1)Immunohistochemistry was used to explore NPRL2 expression in human primary ccRCC clinical specimens;(2)The constructed recombinant expressing plasmids pEGFP-N1-NPRL2 was transfected into primarily cultured cells of human renal cell carcinoma by HG_TransGene;(3)Laser scanning confocal microscopy(LSCM) was used to observe the transfection of the plasmid;(4)qRT-PCR and Western blot were used to detect the mRNA and fusion protein expression of NPRL2 in the transfected cells respectively;(5)The proliferation was detected by the MTT assays;(6)The apoptosis and cell cycle were detected by flow cytometry(FCM).Results:(1) In 78 cases of ccRCC and adjacent normal tissues, NPRL2 protein expression in ccRCC tissues was significantly lower than that in adjacent normal tissues (P<0.001). Clinic pathological analysis showed that NPRL2 expression was significantly correlated with histological grade(P=0.044), TNM stage(P=0.025) and lymph node metastasis (P=0.028), but not with patients'age (P=0.892) and gender (P=0.873);(2) Laser scanning confocal microscopy(LSCM) showed that the plasmid was successfully transfected into primarily cultured cells of human renal cell carcinoma;(3)The expression of NPRL2 mRNA was significantly higher in pEGFP-N1-NPRL2 group than that in pEGFP-N1 group and Blank control group by qRT-PCR(P<0.001);(4) Western blot showed that NPRL2 fusion protein was expressed in pEGFP-N1-NPRL2 group but not in pEGFP-N1 group or Blank control group.(5)MTT assay demonstrated that NPRL2 could obviously inhibit renal cancer cell proliferation;(6)Flow cytometric analysis revealed that NPRL2 could induce renal cancer cells apoptosis and arrest the cell cycle in G0/G1 phase.Conclusion:Decreased expression of NPRL2 is associated with unfavourable pathological, proliferation and apoptotic features in clear cell renal cell carcinoma cells.