AIM: To investigate the incidence of non-small-bowel abnormalities in patients referred for small bowel capsule endoscopy, this single center study was performed.
AIM:To determine the effects of RNAi-mediated inhibition of the growth hormone receptor(GHR)gene on tumors and colon cancer cells in vivo.METHODS:Construction of a eukaryotic vector for human GHR expression,the pcDNA ...AIM:To determine the effects of RNAi-mediated inhibition of the growth hormone receptor(GHR)gene on tumors and colon cancer cells in vivo.METHODS:Construction of a eukaryotic vector for human GHR expression,the pcDNA 6.2-GW/EmGFPsmall interfering RNAs(siRNAs)-GHR plasmid,was used to inhibit GHR expression.Thirty-six BALB/c nude mice were randomly divided into groups and treated with normal saline(NS),recombinant plasmid(G2),growth hormone(GH),5-fluorouracil(FU),G2+FU or G2+FU+GH.Each nude mouse was subcutaneously inoculated with 1×107human colon cancer SW480 cells;the nude mice were weighed before inoculation and on the 2nd,5th,8th,11th,14thand 17thday after inoculation.All nude mice were sacrificed after 17 d.Each subcutaneous tumor was removed and studied.Tumor volume was measured on the 5th,8th,11th,14thand 17thday after inoculation.The expression of GHR protein in the tumor tissue was detected by Western blotting analysis,and the differences in GHR mRNA expression in the tumor tissue were detected by real-time quantitative reverse transcription-polymerase chain reaction.RESULTS:Compared to the control group,the weights of the inoculated nude mice on the 17thday after inoculation were:G2:21.60±0.71 g,GH:21.64±0.45 g,FU:18.94±0.47 g,FU+G2:19.40±0.60 g,G2+FU+GH:21.04±0.78 g vs NS:20.68±0.66 g,P<0.05;the tumor volumes after the subcutaneous inoculation were:G2:9.71±3.82 mm3,FU:11.54±2.42mm3,FU+G2:11.42±1.11 mm3,G2+FU+GH:10.47±1.02 mm3vs NS:116.81±10.61 mm3,P<0.05.Compared to the GH group,the tumor volumes were significantly decreased in the experimental groups.The GHR protein expression(G2:0.39±0.02,FU:0.40±0.02,FU+G2:0.38±0.01,G2+FU+GH:0.39±0.01 vs NS:0.94±0.02,P<0.05)and the GHR mRNA expression(G2:14.12±0.10,FU:15.15±0.44,FU+G2:16.46±0.27,G2+FU+GH:15.37±0.57 vs NS:12.63±0.14,P<0.05)were significantly decreased and increased,respectively,in the experimental groups.CONCLUSION:Inhibition of GHR in human colon cancer SW480 cells resulted in anti-tumor effects in nude mice.展开更多
AIM:To assess the clinicopathological and biological significance of cripto in human colorectal cancer.METHODS:Real-time reverse-transcription polymerase chain reaction(PCR)was used to examine cripto mRNA levels in pr...AIM:To assess the clinicopathological and biological significance of cripto in human colorectal cancer.METHODS:Real-time reverse-transcription polymerase chain reaction(PCR)was used to examine cripto mRNA levels in primary colon cancer and normal colon tissues as well as normal and metastatic lymph nodes from colon cancers.Human colon cancer LS-174T cells were transfected with cripto small interfering RNA(siRNA),and mRNA and protein levels were evaluated using real-time PCR and western blot analysis,respectively.The growth of cancer cells was evaluated using the MTT assay and colony formation in soft agar.Invasion was examined using a Transwell assay,and the expressions of matrix metalloproteinase(MMP)-7 and MMP-9were determined using western blot assay.RESULTS:Cripto was significantly overexpressed in primary colon cancer and metastatic lymph nodes.Silencing cripto gene expression with cripto siRNA resulted in a significant decrease in colony formation in soft agar in the colon cancer cell line LS-174T.Cripto siRNA treatment decreased the migration and invasion capabilities of the colon cancer cell line LS-174T in vitro.Furthermore,cripto siRNA treatment inhibited the expression of matrix MMP-7 and MMP-9.CONCLUSION:The results provide evidence that cripto siRNA could be an effective approach for the inhibition of cancer cell invasion and migration and thus has potential for use in devising novel preventive and therapeutic strategies for colon cancer metastasis.展开更多
文摘AIM: To investigate the incidence of non-small-bowel abnormalities in patients referred for small bowel capsule endoscopy, this single center study was performed.
文摘AIM:To determine the effects of RNAi-mediated inhibition of the growth hormone receptor(GHR)gene on tumors and colon cancer cells in vivo.METHODS:Construction of a eukaryotic vector for human GHR expression,the pcDNA 6.2-GW/EmGFPsmall interfering RNAs(siRNAs)-GHR plasmid,was used to inhibit GHR expression.Thirty-six BALB/c nude mice were randomly divided into groups and treated with normal saline(NS),recombinant plasmid(G2),growth hormone(GH),5-fluorouracil(FU),G2+FU or G2+FU+GH.Each nude mouse was subcutaneously inoculated with 1×107human colon cancer SW480 cells;the nude mice were weighed before inoculation and on the 2nd,5th,8th,11th,14thand 17thday after inoculation.All nude mice were sacrificed after 17 d.Each subcutaneous tumor was removed and studied.Tumor volume was measured on the 5th,8th,11th,14thand 17thday after inoculation.The expression of GHR protein in the tumor tissue was detected by Western blotting analysis,and the differences in GHR mRNA expression in the tumor tissue were detected by real-time quantitative reverse transcription-polymerase chain reaction.RESULTS:Compared to the control group,the weights of the inoculated nude mice on the 17thday after inoculation were:G2:21.60±0.71 g,GH:21.64±0.45 g,FU:18.94±0.47 g,FU+G2:19.40±0.60 g,G2+FU+GH:21.04±0.78 g vs NS:20.68±0.66 g,P<0.05;the tumor volumes after the subcutaneous inoculation were:G2:9.71±3.82 mm3,FU:11.54±2.42mm3,FU+G2:11.42±1.11 mm3,G2+FU+GH:10.47±1.02 mm3vs NS:116.81±10.61 mm3,P<0.05.Compared to the GH group,the tumor volumes were significantly decreased in the experimental groups.The GHR protein expression(G2:0.39±0.02,FU:0.40±0.02,FU+G2:0.38±0.01,G2+FU+GH:0.39±0.01 vs NS:0.94±0.02,P<0.05)and the GHR mRNA expression(G2:14.12±0.10,FU:15.15±0.44,FU+G2:16.46±0.27,G2+FU+GH:15.37±0.57 vs NS:12.63±0.14,P<0.05)were significantly decreased and increased,respectively,in the experimental groups.CONCLUSION:Inhibition of GHR in human colon cancer SW480 cells resulted in anti-tumor effects in nude mice.
基金Supported by The National Natural Science Fund,No.81250035The Jiangsu Province of 333 Engineering talent fund,No.RA2012103Zhenjiang City Social Development Fund No.SH2011031 and SH2011034
文摘AIM:To assess the clinicopathological and biological significance of cripto in human colorectal cancer.METHODS:Real-time reverse-transcription polymerase chain reaction(PCR)was used to examine cripto mRNA levels in primary colon cancer and normal colon tissues as well as normal and metastatic lymph nodes from colon cancers.Human colon cancer LS-174T cells were transfected with cripto small interfering RNA(siRNA),and mRNA and protein levels were evaluated using real-time PCR and western blot analysis,respectively.The growth of cancer cells was evaluated using the MTT assay and colony formation in soft agar.Invasion was examined using a Transwell assay,and the expressions of matrix metalloproteinase(MMP)-7 and MMP-9were determined using western blot assay.RESULTS:Cripto was significantly overexpressed in primary colon cancer and metastatic lymph nodes.Silencing cripto gene expression with cripto siRNA resulted in a significant decrease in colony formation in soft agar in the colon cancer cell line LS-174T.Cripto siRNA treatment decreased the migration and invasion capabilities of the colon cancer cell line LS-174T in vitro.Furthermore,cripto siRNA treatment inhibited the expression of matrix MMP-7 and MMP-9.CONCLUSION:The results provide evidence that cripto siRNA could be an effective approach for the inhibition of cancer cell invasion and migration and thus has potential for use in devising novel preventive and therapeutic strategies for colon cancer metastasis.
文摘目的:探讨小干扰RNA(small interference RNA,siRNA)沉默人结肠癌HT-29细胞livin表达对HT-29细胞增殖、凋亡和侵袭的影响。方法:合成靶向livin的双链siRNA(livin-siRNA),转染HT-29细胞,RT-PCR及Western blotting检测HT-29细胞中livin mRNA及蛋白的表达,MTT实验检测HT-29细胞的增殖,流式细胞术检测HT-29细胞周期分布及凋亡,细胞侵袭实验检测HT-29细胞侵袭性的变化,caspase-3活性检测试剂盒检测caspase-3活性的变化。结果:Livin-siRNA转染后48 h,与空白组、阴性对照组及脂质体组相比,livin-siRNA转染组HT-29细胞中livin mRNA水平明显下降(0.073±0.007 vs 0.395±0.082、0.423±0.025、0.418±0.032,P<0.05),其蛋白表达也明显下调(0.106±0.003 vs 0.456±0.065、0.473±0.078、0.491±0.045,P<0.05)。转染96 h后,livin-siRNA组HT-29细胞增殖能力明显低于对照组及脂质体组(0.564±0.102 vs0.833±0.127、0.860±0.153,P<0.05),且细胞凋亡率升高[(16.5±2.8)%vs(2.4±0.5)%、(3.7±1.0)%,P<0.05]。侵袭实验显示,livin-siRNA转染后,穿过Matrigel膜的HT-29细胞数量明显少于对照组及脂质体组[(31.3±4.5)vs(101.3±8.6)、(97.4±7.8)个,P<0.05)]。livin-siRNA组HT-29细胞的caspase-3活性低于对照组(0.160±0.023 vs 0.347±0.058,P<0.05)。结论:siRNA沉默livin的表达可抑制HT-29细胞的增殖,诱导细胞凋亡,抑制细胞的侵袭。
