| Objective:(1)To elucidate the expression pattern of PARP6 and Survivin in gastric carcinoma and adjacent normaltissues;(2)To explore the relationship between PARP6 and Survivin in gastric cancer and the correlation between their expression and clinicopathological features of gastric cancer;(3)To determine the effect of PARP6 on the proliferation,migration and invasion of gastric cancer cell line BGC823.Methods:(1)Collected the paraffin-embedded specimens of 65 cases of gastric carcinoma from Guizhou Provincial People's Hospital during September 2016to September 2017;In addition,ten samples of normal tissues adjacent to gastric carcinoma were randomly examined as controls;Adopted immunohistochemistry to detect the expression of PARP6 and Survivin in gastric carcinoma and analyzed the correlation between PARP6 and Survivin expression and the clinicopathological features of gastric carcinoma.(2)The expression of PARP6 and Survivin in normal gastric mucosal epithelial cells(GES-1)and gastric cancer cell lines(BGC823)were analyzed by Western Blot;(3)The stable BGC823 cell lines with higher or lower PARP6 were constructed by lentiviral vector transfection;Then the protein expression of PARP6 and Survivin in overexpressed PARP6 cell lines,shRNA PARP6 cell lines and empty plasmid cell lines(control group)were detected by Western Blot.(4)Cell Counting kit-8(cck-8)assay was used to detect the proliferation activities of overexpressed PARP6 cell lines and shRNA PARP6 cell lines at 24h,48h and 72h(5).The migration and invasion abilities of overexpressed PARP6 cell lines and shRNA PARP6 cell lines were compared by wound healing assay,cell invasion assay and migration assay respectively.Results:(1)Immunohistochemical results showed that the high expression rates of PARP6 and Survivin in gastric cancer tissues were 86.2%(56/65)and 92.3%(60/65),respectively,while they were low expressed in adjacent tissues(P<0.05);?~2 test showed that the expressions of PARP6 and Survivin in gastric carcinoma had no significant correlation with age,gender,metastasis,TNM stage and differentiation(P>0.05);A positive correlation between PARP6 and Survivin in gastric carcinoma was obtained by Spearman correlation analysis(r=0.720,P<0.05).(2)Western blotting showed that the protein expression of PARP6 and Survivin in gastric carcinoma cell line(BGC823)were significantly higher than those in normal gastric mucosal epithelial cells(GES-1),both of which had statistical significance(P<0.05).(3)Compared with the control group,Survivin protein was highly expressed in overexpressed PARP6 cell lines,while low expression was observed in shRNA PARP6 cell lines(P<0.05).(4)Cell proliferation assay showed that the OD value of the overexpressed PARP6 cell lines at 24h,48h and 72h was significantly higher than that of the control group(P<0.05),while that of the shRNA PARP6 cell lines was significantly lower than that of the control group(P<0.05).(5)Cell scratch assay showed that the migration rate of overexpressed PARP6 cell lines was significantly higher than that of the control group(P<0.05),while that of shRNA PARP6 cell lines was significantly lower than that of the control group(P<0.05);Transwell assay and invasion experiment showed that the number of cells passing through the chamber of overexpressed PARP6 cell line was significantly higher than that of the control group(P<0.05),while the number of cells passing through the chamber of shRNA PARP6 cell lines was significantly lower than that of the control group(P<0.05).Conclusions:(1)PARP6 and Survivin were highly expressed in gastric cancer tissues and cell lines,possibly contributing to the occurrence of gastric cancer;(2)PARP6 may promote the proliferation,migration and invasion ability of gastric cancer cells through up-regulating Survivin,and participate in the development of gastric cancer. |
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