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Objective:To investigate whether the overexpression of Oct4B1 gene induces epithelial mesenchymal transition in human colorectal cancer SW480 cells and its possible mechanism.Methods: Experimental group(SW480-Oct4B1):Transfection of SW480 cell lines in colorectal cancer with Oct4B1 overexpression plasmid;Control group(SW480-Oct4B1):negative control plasmid with G418 resistance.Stably transfected cell lines were obtained by G418 culture medium.The two groups were compared with:①Detection of Oct4B1 gene expression in stably transfected cell lines by RT-qPCR;②Scratches and Transwell assays were used to estimate migration and invasion;③Detection of EMT related markers E-cadherin,N-cadherin and Vimentin protein expression by Western blot assay;④Detection of Twist gene and protein expression by RT-PCR and Western blot assays.Results: The transient transfection was confirmed by RT-qPCR and the stable transfected cell lines were obtained from two groups of cells transfected with G418 culture medium.Compared with the control group:①RT-qPCR revealed increased expression of Oct4B1 gene in the experimental group(P<0.01);②Cell migration and invasion were significantly increased(P<0.01);③Epithelial marker:the expression of E-cadherin protein was significantly decreased (P<0.01),interstitial marker:the expression of N-cadherin and Vimentin protein was significantly increased (P<0.01);④Twist mRNA and protein expression were significantly increased(P<0.01).Conclusion: Overexpression of Oct4B1 gene can induce epithelial mesenchymal transition in human colorectal cancer SW480 cells,its molecular mechanism may be related to the promotion of Twist expression.
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Objective:To investigate the expression and its possible role of Oct4B1 subtype of Embryonic stem cell transcription factor Oct4 in colorectal cancer stem cells. Methods: 3D microspheres were cultured by suspension culture to human colorectal cancer cell line SW480 cells. The 3D microspheres and SW480 cells were used as the research objects. Whether 3D microspheres were enriched cancer stem cells,we used the methods of cell differentiation experiments,soft agar cloning experiments,and the expression levels of cancer stem cells markers CD133,CD44 detected by flow cytometry. The expression levels of Oct4B1 mRNA were detected by RT-qPCR. Results:3D microspheres could differentiate into normal cancer cells. Compared with the parental SW480 cells,in vitro colony formation was significantly enhanced(P<0. 01),the percentage of positive cells of CD133 and CD44 were significantly increased ( P < 0. 01 ), the expression levels of Oct4B1 mRNA were obviously higher ( P < 0. 01 ) in 3D microspheres. Conclusion: Oct4B1 subtype of Embryonic stem cell transcription factor Oct4 in 3D microspheres enriched human colorectal cancer stem cells,which may be involved in the regulation of colorectal cancer stem cells.
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Objective:To investigate the reversal multidrug resistance effects of curcumin on human colorectal cancer cell lines resistant to oxaliplatin( SW620/OxR) and whether its mechanism was involved in downregulation of STAT3 signaling.Methods: The IC50 value(50%cell growth inhibitory concentrations) of curcumin to SW620/OxR cell lines was obtained by WST-1 reagent,which was one kind of cell proliferation assay.Later experiments included in the experimental group and the control group.In the experimental group,SW620/OxR cell lines were exposed to the previous experiment IC50 concentrations of curcumin plus 2 μmol/L oxaliplatin for 48 h,while in the control group,SW620/OxR cell lines were cultured in medium containing in 2μmol/L oxaliplatin.In the two groups:apoptosis was detected by flow cytometry;the protein expression levels of phosphorylated STAT3(P-STAT3) and P-gp were disclosed by Western blot.Results:The IC50 value of curcumin to SW620/OxR cell lines was 18.9 μmol/L.The apoptosis rate of cells in the control group and the experimental group were respectively ( 5.08 ±1.82 )% and ( 30.69 ±2.94 )%, the apoptosis rate of the experimental group was significantly higher than that of the control group ( P<0.05 ) .The protein expression levels of phosphorylated STAT3(P-STAT3) and P-gp in the experimental group was significantly lower than that in the control group(P<0.05).Conclusion:Curcumin can reverse drug resistance in colorectal cancer cell lines resistant to oxaliplatin, its effect may be associated with downregulation of STAT3 signaling pathways.
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Objective To obtain differential expression genes from colorectal cancer cells derived from colo205 for further research. Methods RNA from colo205 cells,CD133+cells and CD133-cells were sequenced and analyzed by bioinformatics software. Results One hundred and twenty four differential expression genes were obtained, which involves 32 metabolic pathways. Conclusions Large quantities of differential genes can be found among different groups of cells derived from colo205 cells , which can provide epigenetic evidence for colorectal cancer research.
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Objective:To study the clinical application and postoperative complication of double stapling technique (DST) in anus-preserving operation of low and middle rectal cancer. Method:From April 2002 to April 2006, clinical data of 75 cases with low and middle rectal cancer were analyzed retrospectively. Of those 32 cases used DST underwent sphincter-reserving operation(SPO),regard as study group;and 43 cases used SST received SPO, as control group. Clinical pathologic parameters、 postoperative complication and tumor recurrence were compared between the two groups. Results:Operation of all the 75 patients was successful without operative mortality. In DST group patients, defecation function was different from SST group after postoperative 3 and 6 months (P
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Objective:To evaluate the changes of urinary and sexual function after pelvic autonomic nerve preservation with total mesorectal excision operation in patients of rectal cancer . Methods:Fifty-two patients of Dukes B stage received The PANP following TME with follow-up was complete in 38 cases while TME in 35 cases of Dukes B from January 2004 to August 2006. The urinary and sexual dysfunctions of patients in case and control group were analyzed retrospectively by case-control study. Urinary and sexual function were evaluated by means of questionnaires before and after operation.Results:The micturation dysfunction rate was 15.7%(6/38) and 40%(14/35) ,the sexual dysfunction rate was 18.4%(7/38) and 51.4%(18/35)in case and control group respectively,with the difference being significant(P=0.013,P=0.002).Conclusion:The PANP following TME for rectal cancer could reduce sexual and micturition disorder rate and improve the quality of life of the patients after operation.