ABSTRACT
Objective To establish a liquichip method for detecting 6 sub-genotypes of hepatitis C virus(HCV),including 1a, 1b,2a,3a,3b and 6a.Methods The coupling method of PCR amplification and nucleic acid probe was established.The PCR product and the microspheres mixture of the coupled nucleic acid probe were hybridized for establishing the liquichip detection method.The sensitivity and specificity of the established liquichip detection method were evaluated.Nucleic acid in 93 serum samples was detec-ted by this method..Results The established HCV nuclei acid liquichip genotype detection method had the higher specificity and sensitivity,which could detect and classfy 6 HCV sub-genotypes.The sensitivity for HCV 1a,3a and 6a sub-genotypes was 1× 105 copies/PCR;the sensitivity for HCV 1b,2a and 3b sub-genotypes was 1×104 copies/PCR.The detection results in 93 serum samples showed that the this genotyping method had the characteristics of high throughput,rapidness,sentsitivity and specificity. Conclusion This method can be used for the simultaneous and quick detection of 6 HCV sub-genotypes and provides a new meth-od for the genotyping detection of HCV.
ABSTRACT
Objective To analyze the clinical value of microRNAs applied in the early diagnosis of pancreatic cancer.Methods The expression of mir-155,mir-196a,mir-181a,mir-181b in three pancreatic cancer cell lines PANC-1,PaCa-2,AsPC-1 pancreatic cancer,normal pancreatic cells and plasmas of 100 patients with pancreatic cancer ,100 normal volunteers were detected by Real-time PCR,and the results were analyzed.The expression of mir-155,mir-196a,mir-181a,mir-181b in 100 pancreatic cancer tissues and 100 normal pancreatic tissues were detected by immunohistochemical,and the results were analyzed. Results Real-timePCR test results showed that compared with normal pancreatic cells,mir-155,mir-196a expression in three pancreatic cancer cells PANC-1,PaCa-2,AsPC-1 were significantly increased(P<0.05).Compared with healthy volunteers,mir-155,mir-196a,mir-181a,mir-181b expression of plasma in pancreatic cancer patients were significantly higher(P<0.05).Immunohistochemistry results showed that mir-155,mir-196a mir-181a and mir-181b expression in pancreatic tissue were significantly higher than that in normal pancreatic tissue (P <0.05 ).Conclusion The expression of mir-155,mir-196a,mir-181a and mir-181b are high in pancreatic cancer tissues and pancreatic cancer cell lines,which are closely related to the development of pancreatic cancer.Those microRNAs are expected to be cancer markers for early diagnosis of,pancreatic cancer,have highly diagnostic value and broad clinical application.
ABSTRACT
Objective To study the effects of Forkhead box protein M1 (FoxM1) down regulation by small interfering RNA(siRNA) on chemosensitivity and mechanism of human pancreatic cancer cell and its mechanism.Methods Three FoxM1 siRNAs were designed and constructed.All cancer cells were divided into different groups,after transfected with FoxM1 siRNA for different time,the cultured cells were harvested to carry on the next tests.Expression of FoxM1 were determined by red-time PCR and Western blot,and prolifearion and chemosensitivity were evaluated by MTT assay,and the phosphorylation of Akt protein was examined by Western blot.Results FoxM1 siRNA could down-regulate the FoxM1 expression in a dose-and time-dependent manner.The MTF results showed that the inhibit rates was 17.78％,17.56％,35.39％,52.81％,70.98％ indifferentgroups [Con-A + Gemcitabine,Con-B + Gemcitabine,siRNA (3.125nM) + Gemcitabine,siRNA (6.25nM) + Gemcitabine and siRNA(12.5nM) + Gemcitabine,respectively.The phosphorylation of Akt protein was inhibited in a dose-dependent manner.Conclusions FoxM1 siRNA could sensitize human pancreaticr cancer cells chemotherapy sensitivity,it is the one of the important mechanisms through down-regulate Akt phosphorylated levels,but the molecular mechanism need to be explored further.