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Objective To investigate the expression and significance of adaptor protein containing PH domain,PTB domain,and leucine zipper motif 1 (APPL1) in gastric cancers.Methods Expressions of APPL1 protein and mRNA were detected with immunohistochemistry and reverse transcription-polymerase chain reaction (RT-PCR) in 47 surgical specimens of gastric carcinomas (GC) and 27 normal gastric tissue specimens,respectively.Results The expression rate of APPL1 protein was 59.6％ (28/47) in GCs,and 26.1％ (6/23) in normal gastric tissues,with a statistically significant difference between two groups (P ＜0.05).The relative expression of mRNA was 0.821 ±0.141 in GCs,and 0.731 ±0.112 in normal gastric tissues,with a statistically significant difference between two groups (P ＜ 0.05).Conclusions The expressions of APPL1 protein and mRNA are increased in GCs,with a close relationship between GC and APPL1.
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Objective To investigate the influence of c-myc on the growth, proliferation, apoptosis,invasion and cell cycle of the gastric line HFE145. Methods The cDNA of c-myc was subcloned into a constitutive vector pcDNA3.1 followed by transfection in HFE145 by using liposome. Then stable expression clones (HFE-myc) were selected. The apoptosis and cell cycles were detected using flow cytometry. The growth and proliferation were analyzed by making cell growth curves and colony formation assay respectively. The ability of invasion were tested using cell migration assay. Results HFE-myc group grew faster than HFE145and HFE-pc. The cell counts of HFE-myc in five of seven days were more than those of others significantly (P＜0.05). There was no difference between the two control group. Cell cycle analysis showed that HFE-myc group proliferated faster, mean proportion of cells in G2-M was about 25 % and higher significantly (P ＜0.05)than those of the two control groups. Results of colony formation assay showed that the mean colony formation rate of HFE-myc was 0.27 and higher than those of the control groups(P ＜0.05). The results of cell migration assay suggested that the cell migration rate of HFE-myc was not higher significantly than those of the control groups (P ＞0.05). Conclusion c-myc can promote the growth, proliferation. It can increase the proportion of cells in division stage, so promote the division. But it have little influence on the invasion of cells.
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BACKGROUND:Transplantation of autologous stem cells for treatment of liver cirrhosis has been widely reported.But up to now,there exist some concerns for clinical physicians,including relationship between stem cells and post-transplantation prognosis/turnover of liver cirrhosis,directed differentiation of stem cells in the impaired liver,and malignant phenotype.OBJECTIVE:To dynamically monitor the serum levels of alpha-fetoprotein(AFP)and AFP variants(AFP-L3)in decompensated cirrhosis patients following intrahepatic transplantation of peripheral blood stem calls via portal vein and evaluate the safety of this treatment method.METHODS:A total of 44 decompensated cirrhosis patients who underwent intrahepatic transplantation of peripheral blood stem cells via portal vein in the 309 Hospital of Chinese PLA in April 2007 were included in this study.Prior to and after surgery,serum levels of AFP and AFP-L3 were detected by chemiluminescence.Through the use of a positive criterion for liver cirrhosis,i.e.,the proportion of AFP-L3 in AFP[AFP-L3(%)]≥10%,and the relationship between decompensated cirrhosis treatment using stem cells transplantation and the malignant phenotype of liver cancer were analyzed.RESULTS AND CONCLUSION:At 2 months after surgery,serum level of AFP showed a transient increase.There was no significant difference in AFP-L3(%)between prior to and after surgery(P>0.05).No significant difference in AFP-L3-positve rate,as well as AFP-L3(%),existed among patients with different serum level of AFP.These findings indicate that clinical symptoms and liver function of decompensated cirrhosis patients recovered to some extent after transplantation of peripheral blood stem cells via portal vein.Results regarding serum level of AFP-L3,a serological marker of liver cancer,did not demonstrate the appearance of malignant biological phenotype.
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Objective To investigate the effect of recombinant plasmid pshRNA-DNMT3b on expression of DNMT3b mRNA and protein and on the proliferation of bladder cancer T24 cells,and research the function of DNMT3b in the process of bladder tumor formation.Methods There were three groups in this study,which are blank controller,HK and pshRNA-DNMT3b(24h,48h,72h),respectively.T24 cells were cultured routinely and transfected by the recombinant plasmids with lipfectamine 2000.The cells were detected by methods of RT-PCR,western blot and MTT.The varying level of DNMT3b mRNA and expression protein,and the conditions of cellular survival rate were observed.Results The recombinant plasmids were successfully transfected into T24 cell lines.The grey valHe of RT-PCR elctrophoretogram was analyzed by the software of Gel-pro analyzer,the rate of blank controller,HK and pshRNA-DNMT3b(24h,48h,72h),was (99.56±1.24)%,(99.12±1.35)%,(75.77±1.42)%,(44.69±1.05)%and(20.52±0.89)%,respectively.The analytical resuit of western blot image was(99.43±1.28)%,(98.90±1.31)%,(67.83±1.02)%,(43.43±1.05)%and(21.92±0.89)%.There was no statistically difference in survival between blank control and HK(P＞0.05).The group of pshRNA-DNMT3b and other two groups had statistical difference only at the 72th hour and the cell inhibitory growth rate only increase 0.45%.Conclusions The recombinant ptasmid pshRNA-DNMT3b can inhibit the expression of mRNA and protein of DNMT3b effectively.However,it has slight function on inhibiting cell proliferation.