الملخص
<p><b>OBJECTIVE</b>To investigate the expressions of prostate stem cell antigen (PSCA) and Claudin-4 in human pancreatic carcinoma and to discuss its role in the ontogenesis of pancreatic cancer.</p><p><b>METHODS</b>Pancreatic carcinoma tissue microarray was constructed, containing 100 cores of 10 normal adult pancreas tissues, 12 chronic pancreatitis tissues, and 78 pancreatic carcinomas. The expressions of PSCA and Claudin-4 were detected using immunohistochemical method and the relationship between PSCA and Claudin-4 and the pancreatic carcinoma was analyzed.</p><p><b>RESULTS</b>The positive expression rates of PSCA and Claudin-4 protein in pancreatic carcinoma were 79. % and 88. % respectively. PSCA and Claudin-4 staining were more intense in malignant cells than in chronic pancreatic tissues and normal adult pancreas tissues. No evidence was found for an association between expressions of PSCA and Claudin-4 and other variables, including gender, age at surgery, and tumor grade.</p><p><b>CONCLUSIONS</b>The expressions of PSCA and Claudin-4 are related to the pancreatic carcinomas. PSCA and Claudin-4 play a role in the development of pancreatic cancer, but PSCA and Claudin-4 are not correlated with the clinical pathology of tumor.</p>
الموضوعات
Adult , Aged , Female , Humans , Male , Middle Aged , Antigens, Neoplasm , Carcinoma , Genetics , Metabolism , Pathology , Claudin-4 , GPI-Linked Proteins , Gene Expression Regulation, Neoplastic , Membrane Glycoproteins , Genetics , Metabolism , Membrane Proteins , Genetics , Metabolism , Neoplasm Proteins , Genetics , Metabolism , Pancreatic Neoplasms , Genetics , Metabolism , Pathology , Up-Regulationالملخص
<p><b>OBJECTIVE</b>To investigate the expression of calcyclin in pancreatic carcinoma and its relation to the patients' prognosis.</p><p><b>METHODS</b>Human pancreatic carcinoma tissue microarray was constructed, which contained 63 cores of 3 normal adult pancreas tissues, 6 chronic pancreatitis tissues, 51 pancreatic carcinoma tissues and 3 islet cell carcinoma tissues. Immunohistochemistry was performed to detect the expression of calcyclin in these tissues, and the relationship between calcyclin and the clinicopatholoical features of pancreatic carcinoma was analyzed.</p><p><b>RESULTS</b>The positivity rate of calcyclin in the pancreatic carcinoma tissue was 76.5% (39/51), and calcyclin staining was more intense in the malignant cells than in the benign cells (P=0.007), suggesting a correlation between calcyclin expression and pancreatic carcinoma. No evidence was found for an association of calcyclin expression with the variables including the patients' gender, age at surgery, and tumor grade. Weak staining for calcyclin was noted in chronic pancreatitis tissues.</p><p><b>CONCLUSION</b>Calcyclin expression is related to the pancreatic carcinomas and up-regulation of calcyclin expression is possibly an early event in pancreatic and pragression of development cancer.</p>
الموضوعات
Adult , Aged , Female , Humans , Male , Middle Aged , Biomarkers, Tumor , Genetics , Metabolism , Cell Cycle Proteins , Genetics , Metabolism , Gene Expression Regulation, Neoplastic , Immunohistochemistry , Pancreatic Neoplasms , Genetics , Metabolism , Pancreatitis, Chronic , Genetics , Metabolism , S100 Calcium Binding Protein A6 , S100 Proteins , Genetics , Metabolismالملخص
<p><b>OBJECTIVE</b>To construct an expression vector of small interfering RNA (siRNA) against survivin and observe its effects on survivin expression and proliferation of human pancreatic cancer cell line PC-2 and breast cancer cell line MCF-7.</p><p><b>METHODS</b>Constructed an expression vector of siRNA against survivin and transfected it into PC-2 and MCF-7 cells using lipofectamine 2000. The expression of survivin was detected by semi-quantitive RT-PCR and immunohistochemistry, and its effects on proliferation of PC-2 and MCF-7 cells were detected by MTT assay.</p><p><b>RESULTS</b>The introduction of sequence-specific siRNA could efficiently suppress survivin expression at both mRNA and protein levels in the two cancer cell lines. In PC-2 cells, the expression inhibition rates were 81.25% at mRNA level and 74.24% at protein level. In MCF-7 cells, the expression inhibition rates were 64.91% at mRNA level and 79.72% at protein level. The proliferation of PC-2 and MCF-7 cells was also suppressed, and 24 and 48 hours after the cells were reseeded, the proliferation inhibition rates of PC-2 cells were 28.00% and 33.38%, and that of MCF-7 cells were 31.58% and 33.02%, respectively.</p><p><b>CONCLUSIONS</b>The expression vector of siRNA against survivin can block survivin expression in PC-2 and MCF-7 cells efficiently and specifically. Down regulation of survivin expression can suppress proliferation of PC-2 and MCF-7 cells. Survivin RNAi may have potential value in gene therapy of human cancers.</p>
الموضوعات
Female , Humans , Base Sequence , Breast Neoplasms , Pathology , Therapeutics , Cell Line, Tumor , Cell Proliferation , Gene Expression , Inhibitor of Apoptosis Proteins , Microtubule-Associated Proteins , Genetics , Metabolism , Pancreatic Neoplasms , Pathology , Therapeutics , Plasmids , Genetics , RNA, Messenger , Genetics , Metabolism , RNA, Neoplasm , Genetics , Metabolism , RNA, Small Interfering , Genetics , Therapeutic Uses , Transfectionالملخص
<p><b>OBJECTIVE</b>Blocking the expression of survivin with RNA interference techniques, the effects of suppressing proliferation and inducing apoptosis of breast cancer MCF-7 cells were investigated.</p><p><b>METHODS</b>A siRNA eukaryotic expression vector against survivin was constructed and transfected into breast cancer MCF-7 cells with lipofectamine 2000. The changes of survivin expression were detected by semi-quantitive RT-PCR and immunohistochemistry. The effect of suppressing proliferation of MCF-7 cell was detected by MTT assay. The effect of inducing MCF-7 cell apoptosis was detected by TUNEL assay.</p><p><b>RESULTS</b>The sequence-specific siRNA can efficiently block the expression of survivin both at mRNA and protein levels. The expression inhibition rate was 64.9% at mRNA level detected by semi-quantitive RT-PCR and 79.7% at protein level detected by immunohistochemistry. Blocking the expression of survivin can suppress proliferation of MCF-7 cells significantly. At 24 and 48 h after the cells were reseeded, the proliferation inhibition rates were 31.6% and 33.0%, respectively. At 24 h after transfection, apoptosis was induced in 12.9% of the cells as detected by TUNEL assay.</p><p><b>CONCLUSION</b>Blocking the expression of survivin with RNA interference technology can significantly suppress proliferation of MCF-7 cells and induce apoptosis to a certain degree. RNAi targeted to survivin has a potential value in gene therapy of breast cancer.</p>
الموضوعات
Female , Humans , Apoptosis , Breast Neoplasms , Metabolism , Pathology , Cell Line, Tumor , Cell Proliferation , Genetic Therapy , Genetic Vectors , Inhibitor of Apoptosis Proteins , Microtubule-Associated Proteins , Genetics , Physiology , Neoplasm Proteins , Genetics , Physiology , RNA Interference , RNA, Messenger , Genetics , RNA, Small Interfering , Genetics , Metabolism , Pharmacology , Transfectionالملخص
<p><b>OBJECTIVE</b>To investigate the effect of a sequence-specific small interfering RNA (siRNA) in suppressing survivin expression and cell proliferation and inducing apoptosis of PC-2 cells.</p><p><b>METHODS</b>The plasmid expression vector of siRNA targeted against survivin was constructed and transfected into PC-2 cells with Lipofectamine 2000. The changes of survivin expression were detected by semi-quantitative RT-PCR and immunohistochemical SP methods. The effect of siRNA in suppressing the proliferation of PC-2 cells was detected by MTT assay, and its role in inducing PC-2 cell apoptosis evaluated by flow cytometry.</p><p><b>RESULTS</b>The sequence-specific siRNA effectively suppressed survivin expression at both mRNA and protein levels with inhibition rate of 81.25% at mRNA level and 74.24% at protein level. Survivin expression suppression significantly inhibited the proliferation of PC-2 cells, and at 24 and 48 h after cell seeding, the proliferation inhibition rate was 28.00% and 33.38% respectively; 24, 48 h after the transfection, apoptosis occurred in 8.46% and 7.53% of the cells, respectively.</p><p><b>CONCLUSIONS</b>The plasmid expression vector for the siRNA against survivin constructed in the study can effectively and specifically suppress survivin expression in PC-2 cells, and blocking survivin expression suppresses PC-2 cell proliferation and induces cell apoptosis. siRNA targeted against survivin has a potential value in gene therapy for pancreatic cancer.</p>
الموضوعات
Humans , Apoptosis , Genetics , Cell Line, Tumor , Cell Proliferation , Cell Survival , Genetics , Inhibitor of Apoptosis Proteins , Microtubule-Associated Proteins , Genetics , Neoplasm Proteins , Genetics , Pancreatic Neoplasms , Genetics , Pathology , RNA Interference , RNA, Messenger , Genetics , RNA, Small Interfering , Genetics , Transfectionالملخص
<p><b>OBJECTIVE</b>To investigate the role of Ras antisense oligoribonucleotide (ASODN) in multidrug resistance (MDR) of pancreatic carcinoma Pc-2 cells.</p><p><b>METHODS</b>Ras and P-gp expression was suppressed by Ras ASODN. Sensitivity of Pc-2 cells to chemotherapy was determined by the MTT assay. MDR-1 mRNA level was detected by fluorogenic probe quantitative reverse transcription polymerase chain (RT-PCR) method. Flow cytometry (FCM) was used to detect the accumulative concentration of adriamycin (ADR) in the cells.</p><p><b>RESULTS</b>Ras ASODN significantly inhibited the Ras and P-gp expression (P < 0.05), increased the sensitivity of Pc-2 cells to chemotherapeutic agents (P < 0.05), decreased MDR-1 gene level in Pc-2 cells (P < 0.05), and increased the intracellular intake of ADR in Pc-2 cells (P < 0.05).</p><p><b>CONCLUSION</b>Ras ASODN may enhance the sensitivity of multidrug-resistant pancreatic cancer Pc-2 cells to chemotherapeutic agents by regulating MDR-1 gene level.</p>