Objective To study the expression and significance of multidrug resistance-associated protein (MRP) gene in hepatocellular carcinoma (HCC). Methods Reverse transcription polymerase chain reaction (RT-PCR) assay was used to detect the expression of MRP mRNA in 25 fresh specimens of the primary HCC and its surrounding liver tissues. Immunohistochemistry LSAB technique was adopted to test MRP in 60 HCC specimens. The drug sensitivity was also tested by flow cytometry.Results The positive expression rates of MRP mRNA and MRP protein in primary HCC were 44.00%(11/25) and 45.00%(27/60) respectively. All the intensity of expression was low, but significant higer than its surrouding liver tissues (P＜0.05). The intensity and expression rate of MRP protein in 5 recurrent HCC had a tendency to increase. There was a correlation between the expression of MRP mRNA and MRP protein in 25 patients using RT-PCR and immunohistochemistry assay (Plt;0.05). Detected by flow cytometry, the average sensitivity of drugs in vitro of 60 HCC sp-cimens were 5-FU （15.80±7.63）%，DDP（18.45±9.59）%，ADM（17.95±7.99）%，MMC（16.60±8.69）% and CTX（17.40±10.14）%. Only 5FU and ADM were significantly affected by the expression of MRP protein (Plt;0.05).Conclusion The expression of MRP in primary HCC may be one of the important mechanisms of the intrinsic and acquired drug resistance in HCC. To study the expression of MRP could give a predictive value in HCC chemotherapy.
We have devised a highly sensitive, specific, and quantitative assay for multidrug resistance (mdr1) mRNA expression based on the reverse transcription-polymerase chain reaction (RT-PCR). mdr1 mRNA levels were detected in 30 human primary hepatocellular carcinoma (PHC) tissue and adjacent liver tissue. Five of the patients had received chemotherapy before hepatectomy. The results show that the level of expression of mdr1 gene is higher in tumor tissue than in adjacent liver tissue. mdr1 gene is overexpressed in PHC after chemotherapy. Furthermore, mdr1 gene expression in the treated tumor adjacent liver tissue is higher than that in untreated tumor adjacent liver tissue. Our results indicated that overexpression of mdr1 gene may be responsible for the intrinsic and acquired drug resistance of PHC.
Objective To investigate the transfection and expression of recombinant plasmid human vascular endothelial growth factor 165/pcDNA3. 1 （hVEGF165/pcDNA3. 1） in myocardial cells, and to build foundation for gene therapy and cell therapy of coronary artery disease （CAD）. Methods Myocardial cells were cultured in vitro and transfected by hVEGF165/pcDNA3.1 with liposome; then transient expressed protein was detected by reverse transcriptase-polymerase chain reaction （RT-PCR）, immunochemistry and Western blotting. Results A strap as hVEGF165 was obtained by RT-PCR, the protein of hVEGF165 was found in myocardial cells by immunochemistry and in supernatant by Western blotting. Conclusion The recombinant plasmid hVEGFI65/pcDNA3. 1 can be expressed in myocardial cells, and may be used in studying CAD by gene therapy and cell transplantation.
Objective To explore the effect of connective tissue growth factor on the pathogenesis of hypertrophic scar and keloid tissue. Methods The content of hydroxyproline was determined and the expression of connective tissue growth factor gene was detected by the reverse transcription-polymerase chain reaction and image analysis technique in 5 normal skins, 15 hypertrophic scars and 7 keloid tissues. Results The contents of hydroxyproline in the hypertrophic scar(84.10±1.76) and keloid tissue (92.38±2.04) were significantly higher than that of normal skin tissue (26.52 ± 4.10) (P lt; 0.01). The index of connective tissue growth factor mRNA in the hypertrophic scar (0.78 ± 0.63) and keloid tissue (0.84 ± 0.04) were higher than that of normal skin tissue ( 0.09 ± 0.25) (P lt; 0.01). Conclusion Connective tissue growth factor may play an important role in promoting the fibrotic process of hypertrophic scar and keloid tissue.
【摘要】 目的 研究过氧化酶增殖因子活化受体γ（peroxisome proliferator-activated receptor gamma， PPARγ）和血管内皮生长因子C（vascular endothelial growth factor C， VEGF-C）在胃癌组织中表达的相关性，分析其表达与临床病理因素之间的关系。 方法 2005年3月-2009年3月间采用逆转录-聚合酶链反应方法检测36例胃癌手术标本中PPARγ和VEGF-C mRNA的表达，同时选取相同患者的胃正常组织作为对照。 结果 PPARγ mRNA在胃癌中的表达量高于癌旁正常组织，两者的差异有统计学意义（P=0.007）；VEGF-C在胃癌中的表达量高于癌旁正常组织，两者的差异有统计学意义（P=0.004）；PPARγ的表达与VEGF-C表达无关联性（r=0.135，P=0.414）；PPARγ表达与胃癌组织中浸润程度有关（χ2=4.620，P=0.032）、淋巴结转移有关（χ2=15.753，P=0.000）和临床病理分期有关（χ2=4.610，P=0.032）；VEGF-C表达与胃癌组织中淋巴结转移有关（χ2=4.729，P=0.030）、远处转移有关（χ2=4.064，P=0.044）和临床病理分期有关（χ2=6.300，P=0.012）。 结论 PPARγ和VEGF-C可能在胃癌新生淋巴管形成中起重要作用，两者的表达水平与胃癌患者的病情判断及预后评价密切相关。【Abstract】 Objective To investigate the significance of expressions of peroxisome proliferator-activated receptor gamma (PPARγ) and vascular endothelial growth factor C (VEGF-C) in gastric carcinoma as well as their correlation, and to study the relationship between the expressions and clinicopathologic characteristics of gastric carcinoma. Methods Thirty-six pairs of normal mucosa and cancer specimens were obtained from patients who had undergone gastric operation for primary gastric carcinoma and subjected to reverse transcription-ploymerase chain reaction (RT-PCR) for PPARγ and VEGF-C mRNA detection. Results The positive rate and level of PPARγ mRNA expression were higher in gastric cancer tissues than those in normal gastric mucosa (P=0.007). The positive rate and level of VEGF-C mRNA expression were also higher in gastric cancer tissues than those in normal gastric mucosa (P=0.004). Simultaneously, the expression of PPARγ was not correlated with that of VEGF-C (r=0.135, P=0.414), while the highly productions of PPARγ and VEGF-C in gastric carcinomas were both associated with the lymph node metastasis and the clinical stage (Plt;0.05). Conclusion PPARγ and VEGF-C may play an important role in the lymphangio-genesis of gastric cancer, and united detection of PPARγ and VEGF-C expressions may be correlated with making diagnosis, evaluating prognosis in patients with gastric cancer at the same time.
Objective To study the significance of c-met mRNA in axillary drainage after operations for breast cancer. Methods RT-PCR assay was used to examine c-met mRNA in axillary drainage after operations in 52 cases of breast cancer. The relationships between the expression of c-met and the tumor size, metastatic lymph nodes， the expressions of estrogen receptor (ER), progesterone receptor (PR) and c-erbB-2 were analyzed, respectively. In addition, the effect of douching operative field with 5-FU and distilled water on the expression of c-met mRNA was also analyzed. Results ①The proto-oncogene c-met mRNA could be detected in axillary drainage after operations for breast cancer by RT-PCR, and its positive rate was higher than that in routine pathological detection for micrometastasis in the axillary lymph nodes (P＜0.05). ②The expression of c-met mRNA was correlated with both the metastatic lymph nodes and tumor size. ③There was no significant relationship between the expression of c-met mRNA and the expressions of ER, PR and c-erbB-2. ④Dounching operative field with 5-FU and distilled water could decrease the expression of c-met mRNA.Conclusion The proto-oncogene c-met mRNA may be an ideal and specific marker for dectecting micrometastasis of breast cancer. In addition, it also suggests that the examination of c-met mRNA in the axillary drainage by RT-PCR assay could detect the micrometastasis in axillary lymph nodes much easier and more accurately than routine pathological method.
【Abstract】Objective To investigate the expression of the mRNA of cancer-testis antigen 9 (CT9) gene in hepatocellular carcinoma. Methods The expression of CT9 mRNA was detected through RT-PCR in HCC tissues and their adjacent non-HCC tissues from 45 HCC patients. From CT9 RT-PCR positive products, 3 samples were selected randomly and were sequenced. ResultsCT9 mRNA was detectable in 51.1%(23/45) of HCC samples, and no expression of CT9 mRNA was detected in the adjacent non-HCC tissues. In addition, the RTPCR products were proved to be CT9 cDNA by DNA sequencing. No relationship was found between the expression of CT9 mRNA and clinical factors such as age, sex, tumor size, degree of tumor differentiation, serum αfetoprotein level and infection of hepatitis B virus or hepatitis C virus （Pgt;0.05）. ConclusionCT9 mRNA is expressed with high percentage and specificity in hepatocellular carcinomas. The CT9 gene product is a potential target for antigenspecific immunotherapy of HCC.
【Abstract】ObjectiveTo discuss the possibility and clinical significance of SSX-1 mRNA used as specific marker for examining HCC cells in peripheral blood of HCC patients. MethodsUsing the RT-PCR method, the SSX1 mRNA of the peripheral blood was examined in 25 cases of HCC patients and 20 non-HCC patients. The same method was used to detect the expression of SSX-1 mRNA in the tumor tissues, para-tumor tissues, cirrhosis tissues and normal hepatic tissues. A randomized sample was extracted for DNA sequencing from positive electrophoresis expression samples of SSX-1 to examine the reliability of results. ResultsThe expression rates of SSX-1 mRNA were 60%(15/25) and 40%（10/25） respectively in tumor tissues of HCC and the corresponding peripheral blood. SSX-1 mRNA were not expressed in para-tumor tissues,cirrhosis tissues and normal hepatic tissues. The DNA sequence -confirmed that the RTPCR products were true target cDNA. No relationships were found between the expression of SSX-1 gene and clinical characteristics, such as age, sex, tumor size, TNM stage， extent of differentiation and serum AFP level (Pgt;0.05). However, in 33%（3/9） patients with normal serum AFP (lt;20 μg/L), specific expression of SSX1 mRNA was observed. ConclusionHigh specific expression of SSX-1 mRNA is observed in the peripheral blood of patients with HCC, it suggests that applying it as a tumor marker to detect HCC cells in peripheral blood is an adjuvant diagnostic tool. The expression of SSX-1 mRNA in the peripheral blood is observed in the group HCC patients whose serum AFP (lt;20 μg/L) are normal, which suggests that applying both SSX-1 mRNA and AFP as tumor markers together might be useful to improve the diagnostic accuracy for HCC.