This study examined the expression of cell adhesion molecule 1 （CADM1） in pancreatic cancer and the possible mechanism. The expression of CADM 1 was detected by immunohistochemistry in tissues of pancreatic cancer, ...This study examined the expression of cell adhesion molecule 1 （CADM1） in pancreatic cancer and the possible mechanism. The expression of CADM 1 was detected by immunohistochemistry in tissues of pancreatic cancer, pancreatitis, and normal pancreas. The plasmid pcDNA3.1-Hy- gro（＋）/CADM1 was transfected into PANC-1 cells （a pancreatic cancer cell line）. The expression of CADM1 in the transfected cells was determined by RT-PCR and Western blotting. Cell growth was measured by the MTT method and cell apoptosis by flow cytometry. The results showed that CADM1 was weakly expressed in tissues of pancreatic cancer in contrast to its high expression in normal pancreatic and pancreatitis tissues. The expression level of CADM in pancreatic caner was intensely correlated with the differentiation degree, lymph node metastasis and TNM stages. The growth of CADMl-transfected PANC-1 cells was significantly suppressed in vitro by a G1 cell cycle arrest and apoptosis occurrence. It was concluded that re-expression of CADM1 inhibits the growth of pancreatic cancer cells and induces their apoptosis in vitro. As a tumor suppressor gene, CADM1 plays an important role in the occurrence, progression and metastasis of pancreatic cancer.展开更多
OBJECTIVE To construct a eukaryotic expression vector for RNA interference of the human cyclinD1 gene, and to detect its interference effect in human ovarian cancer cells （HO-8910）. METHODS Four target gene segments...OBJECTIVE To construct a eukaryotic expression vector for RNA interference of the human cyclinD1 gene, and to detect its interference effect in human ovarian cancer cells （HO-8910）. METHODS Four target gene segments were synthesized and cloned into the pSUPER vector respectively to construct four recombinant eukaryotic expression vectors, pSUPER-C1-4. The four recombinant vectors were identified by enzyme digestion analysis and DNA sequencing. Then HO-8910 cells were transfected with the pSUPER-C1-4 vectors and subjected to G418 selection. In G418-resistant cells, the interference effect was detected by RT-PCR. RESULTS Enzyme digestion analysis and DNA sequencing showed that the target segments were cloned into the pSUPER vector. The four recombinant vectors inhibited transcription of the cyclinD1 gene. The pSUPER-C2 vector had a better interference effect. CONCLUSION The sequence-specific siRNA effectively interfered with expression of the cyclinD1 gene that was selected. The transcription and expression of the cyclinD1 gene were inhibited effectively by the constructed RNAi eukaryotic expression vectors in the ovarian cancer cells. These results indicate that it is possible to search for a new tumor gene therapy method,展开更多
Background and Objective Lung cancer is the leading cause of cancer death in both men and women in both China and worldwide. Apoptosis is a highly regulated,
目的:初步探讨Bmi-1基因对胶质瘤细胞凋亡状况的影响。方法:采用siRNA技术干扰沉默U251胶质瘤细胞中Bmi-1基因,RT-PCR法检测干扰效果,流式细胞仪检测U251细胞的凋亡状况,one way ANOVA和t检验进行统计学分析。结果:RT-PCR显示U251细胞RN...目的:初步探讨Bmi-1基因对胶质瘤细胞凋亡状况的影响。方法:采用siRNA技术干扰沉默U251胶质瘤细胞中Bmi-1基因,RT-PCR法检测干扰效果,流式细胞仪检测U251细胞的凋亡状况,one way ANOVA和t检验进行统计学分析。结果:RT-PCR显示U251细胞RNA干扰组Bmi-1 mRNA凝胶电泳条带呈弱阳性,阴性对照组和未处理组Bmi-1 mRNA条带呈强阳性,灰度比统计分析,差异具有统计学意义,siRNA可以明显降低U251细胞中Bmi-1基因的表达;流式细胞仪显示干扰组U251细胞的凋亡率为(32.53±6.33)%,高于阴性对照组的凋亡率(21.91±5.63)%,统计分析其差异具有统计学意义。结论:Bmi-1基因可以抑制胶质瘤细胞的凋亡,有可能促进了胶质细胞瘤的发生、发展。展开更多
文摘This study examined the expression of cell adhesion molecule 1 （CADM1） in pancreatic cancer and the possible mechanism. The expression of CADM 1 was detected by immunohistochemistry in tissues of pancreatic cancer, pancreatitis, and normal pancreas. The plasmid pcDNA3.1-Hy- gro（＋）/CADM1 was transfected into PANC-1 cells （a pancreatic cancer cell line）. The expression of CADM1 in the transfected cells was determined by RT-PCR and Western blotting. Cell growth was measured by the MTT method and cell apoptosis by flow cytometry. The results showed that CADM1 was weakly expressed in tissues of pancreatic cancer in contrast to its high expression in normal pancreatic and pancreatitis tissues. The expression level of CADM in pancreatic caner was intensely correlated with the differentiation degree, lymph node metastasis and TNM stages. The growth of CADMl-transfected PANC-1 cells was significantly suppressed in vitro by a G1 cell cycle arrest and apoptosis occurrence. It was concluded that re-expression of CADM1 inhibits the growth of pancreatic cancer cells and induces their apoptosis in vitro. As a tumor suppressor gene, CADM1 plays an important role in the occurrence, progression and metastasis of pancreatic cancer.
文摘OBJECTIVE To construct a eukaryotic expression vector for RNA interference of the human cyclinD1 gene, and to detect its interference effect in human ovarian cancer cells （HO-8910）. METHODS Four target gene segments were synthesized and cloned into the pSUPER vector respectively to construct four recombinant eukaryotic expression vectors, pSUPER-C1-4. The four recombinant vectors were identified by enzyme digestion analysis and DNA sequencing. Then HO-8910 cells were transfected with the pSUPER-C1-4 vectors and subjected to G418 selection. In G418-resistant cells, the interference effect was detected by RT-PCR. RESULTS Enzyme digestion analysis and DNA sequencing showed that the target segments were cloned into the pSUPER vector. The four recombinant vectors inhibited transcription of the cyclinD1 gene. The pSUPER-C2 vector had a better interference effect. CONCLUSION The sequence-specific siRNA effectively interfered with expression of the cyclinD1 gene that was selected. The transcription and expression of the cyclinD1 gene were inhibited effectively by the constructed RNAi eukaryotic expression vectors in the ovarian cancer cells. These results indicate that it is possible to search for a new tumor gene therapy method,
基金supported by the following grants to Qinghua ZHOU:The Key Project of National Natural Science Foundation of China (30430300)The Major Project of Tianjin Sci-Tech Support Programme (07SYSYS F05000)+3 种基金The Key Project of Tianjin Sci-Tech Support Programme (06YFS ZSF05300)The Building Project of Tianjin Sci-Tech Innovation Platform (07SYSYJC27900)863 National Major Projects (2006AA02A401)National Key Basic Research and Development Plan (973 Plan) program (20 07CBS914800)
文摘Background and Objective Lung cancer is the leading cause of cancer death in both men and women in both China and worldwide. Apoptosis is a highly regulated,
文摘目的:初步探讨Bmi-1基因对胶质瘤细胞凋亡状况的影响。方法:采用siRNA技术干扰沉默U251胶质瘤细胞中Bmi-1基因,RT-PCR法检测干扰效果,流式细胞仪检测U251细胞的凋亡状况,one way ANOVA和t检验进行统计学分析。结果:RT-PCR显示U251细胞RNA干扰组Bmi-1 mRNA凝胶电泳条带呈弱阳性,阴性对照组和未处理组Bmi-1 mRNA条带呈强阳性,灰度比统计分析,差异具有统计学意义,siRNA可以明显降低U251细胞中Bmi-1基因的表达;流式细胞仪显示干扰组U251细胞的凋亡率为(32.53±6.33)%,高于阴性对照组的凋亡率(21.91±5.63)%,统计分析其差异具有统计学意义。结论:Bmi-1基因可以抑制胶质瘤细胞的凋亡,有可能促进了胶质细胞瘤的发生、发展。
