Influences of pancreatic cancer cell PANC-1 proliferation of downregulating Berlin1 expression / 中国综合临床

ABSTRACT

Objective To observe the influences of Bcl-2 expression and pancreatic cancer cell proliferation after downregulating Beclin1 expression and discuss the mechanism.Methods Target interfering plasmid siBeclin1 was constructed and transfected to PANC-1 cell,and cells were divided into non transfected group(Beclin1 group),blank control group(control siNegative group) and transfection group(siBeclin1 group).Western blot and qRT-PCR were used to detect the transfection efficiency and Bcl-2 expression after transfection.Proliferation was detected by MTT.Results Western blot and qRT-PCR results showed that expression of Beclin1 in siBeclin1 group was lower than Beclin1 and siNegative control groups (27.823± 1.432,8.635±1.481,26.904±1.098;F=8.176,P＜0.01),qRT-PCR showed that the Beclin1 mRNA level of PANC-1 cells in siBeclin1 group was lower than Belcin1 and siNegative groups (0.421 ±0.157,0.194±0.104,0.399 ±0.123;F=5.239,P＜0.01),and the silencing rate was about 60％-70％.Bcl-2 protein expression and levels of mRNA in siBeclin1 group were significantly higher than that of Beclin1 group(26.912±1.927,8.004±1.534,t =7.329,P＜0.01;0.582±0.297,0.217±0.186,t =6.835,P＜0.01);MTT results showed that the proliferation of PANC-1 cells were strengthened,(61.54±6.81)％ and (46.78±7.28)％ at 72 h,(76.39±7.26)％ and (54.27±8.17)％ at 96 h,and the difference was significant(t=3.674,P＜0.05;t =10.185,P ＜0.01).Conclusion Proliferation of pancreatic cancer cells are strengthened after Beclin1 expression downregulating,so Beclin1 may inhibit the proliferation of pancreatic cancer cells by Bcl-2 downregulating.Beclin1 can be a target candidate gene of pancreatic cancer therapy.