Objective To investigate the inhibitory effects of RNA interference (RNAi) expression vector on the expression of survivin in pancreatic cancer cell PANC-1. Methods The protein and mRNA expressions of survivin were examined with immunofluorescence and RT-PCR. The survivin gene was cloned into the T-vector and sequenced. The RNAi expression vectors targeting survivin, named si-svv-1 and si-svv-2 respectively according to whether they harbored a mutation or no mutation, were constructed and transfected into PANC-1 cells with liposome. The expression of survivin mRNA was detected with RT-PCR. Apoptosis of PANC-1 cells was analyzed with DNA ladder and FACS. Results There was a high degree expression of survivin in PANC-1 cells. The expression of survivin was not inhibited by RNAi expression vectors si-svv-1, but inhibited about (72.43±8.04)% by si-svv-2 and the apoptosis rate of PANC-1 cells increased to (12.36±1.44)% after 72 h. Conclusion The RNAi expression vector can effectively inhibit the expression of survivin in pancreatic cancer cell PANC-1 cells and induce the apoptosis in PANC-1 cells.
Objective To investigate the frequency of variant at XmnⅠ, MspⅠ sites of apolipoprotein (Apo) AⅠ-CⅢ-AⅣ gene cluster and its relation to cholesterol cholecystolithiasis in Chinese population. Methods The restriction fragment length polymorphisms (RFLP) at Xmn Ⅰ, MspⅠ sites of ApoAⅠ-CⅢ-AⅣ gene cluster was studied by using polymerase chain reaction (PCR) in healthy subjects (control group) and patients with gallstones(gallstone group) from a population of Chinese with the Han nationality in Sichuan Province. Results The X1, M1 alleles were the major alleles in both gallstone group and control group, and X1X1, M1M homozygous genotypes were the most frequent ones. The frequencies of X1X2, X2X2 and X2 alleles in female patients of the gallstone group were significantly higher than those in female of control group (P<0.05), and the frequencies of X1X1 and X1 alleles in control group were significantly higher than those in gallstone group (P<0.05). The frequencies of M1M1 in male patients of the gallstone group were significantly lower than those in male of control group (P<0.05), whereas, the frequencies of M1M2 were significantly higher in gallstone group (P<0.05). Conclusion The polymorphism of XmnⅠ RFLP of ApoAⅠ-CⅢ-AⅣ gene cluster in female and the polymorphism of MspⅠ RFLP of ApoAⅠ-CⅢ-AⅣ gene cluster in male may be associated with cholesterol cholecystolithiasis in Chinese population.