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Effect of Tumor Suppressor Gene NPRL2 on Proliferation and Apoptosis of Human Renal Cancer Cell Line 786-O
Jiang Li, Tang Wei*
Department of Urinary Surgery, First Affiliated Hospital, Chongqing Medical University, Chongqing 400016, China
Abstract: The tumor suppressor gene NPRL2 expressed obviously in many normal human tissues， but
reduced in expression in many human tumors significantly. In this experiment， the recombinant expressing plasmids
pEGFP-N1-NPRL2 were constructed and then transfected into 786-O cells by HG_TransGene. Fluorescent
microscopy， RT-PCR and Western blot were used to detect the mRNA and protein expression of NPRL2 in the
transfected cells， respectively. The proliferation， apoptosis and cell cycle of 786-O cells were detected by the
MTT assays and flowcytometry. The results showed that after transfected into 786-O cells， both the mRNA and
protein levels of NPRL2 were increased in pEGFP-N1-NPRL2 group compared with pEGFP-N1 group and control
group (P<0.05). MTT assay demonstrated that the value of D490 of the pEGFP-N1-NPRL2 group， pEGFP-N1
group and control group were 0.654±0.030， 1.528±0.022 and 1.572±0.036， respectively. Compared with the other
two groups， pEGFP-N1-NPRL2-infected 786-O cells showed a significant decrease in proliferation (P<0.05). The
cell cycle assay indicated that the apoptotic rate of the pEGFP-N1-NPRL2 group， pEGFP-N1 group and control group were 18.82%±0.40%， 5.65%±0.12% and 5.85%±0.07%， and the proportion of the cell at G0/G1 phase was
69.80%±1.40%， 46.24%±1.30% and 47.03%±0.45%. The cells at G1 phase and apoptotic rate were both increased
(P<0.05) in recombinant group. The cell cycle was arrested in the G0/G1 phase after transfection compared with the
other two groups. The results suggested that the transfection of NPRL2 gene could suppress the proliferation， induce
cell apoptosis and arrest cell cycle at G0/G1 phase of 786-O cells.
reduced in expression in many human tumors significantly. In this experiment， the recombinant expressing plasmids
pEGFP-N1-NPRL2 were constructed and then transfected into 786-O cells by HG_TransGene. Fluorescent
microscopy， RT-PCR and Western blot were used to detect the mRNA and protein expression of NPRL2 in the
transfected cells， respectively. The proliferation， apoptosis and cell cycle of 786-O cells were detected by the
MTT assays and flowcytometry. The results showed that after transfected into 786-O cells， both the mRNA and
protein levels of NPRL2 were increased in pEGFP-N1-NPRL2 group compared with pEGFP-N1 group and control
group (P<0.05). MTT assay demonstrated that the value of D490 of the pEGFP-N1-NPRL2 group， pEGFP-N1
group and control group were 0.654±0.030， 1.528±0.022 and 1.572±0.036， respectively. Compared with the other
two groups， pEGFP-N1-NPRL2-infected 786-O cells showed a significant decrease in proliferation (P<0.05). The
cell cycle assay indicated that the apoptotic rate of the pEGFP-N1-NPRL2 group， pEGFP-N1 group and control group were 18.82%±0.40%， 5.65%±0.12% and 5.85%±0.07%， and the proportion of the cell at G0/G1 phase was
69.80%±1.40%， 46.24%±1.30% and 47.03%±0.45%. The cells at G1 phase and apoptotic rate were both increased
(P<0.05) in recombinant group. The cell cycle was arrested in the G0/G1 phase after transfection compared with the
other two groups. The results suggested that the transfection of NPRL2 gene could suppress the proliferation， induce
cell apoptosis and arrest cell cycle at G0/G1 phase of 786-O cells.
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