Detection of tumor cells in peripheral blood of patients with gastric cancer using mRNA of MAGE genes as markers / 中华胃肠外科杂志

<p><b>OBJECTIVE</b>To develop a sensitive and specific RT-PCR assay using the mRNA of MAGE-1 and MAGE-3 genes as specific tumor markers for the detection of the tumor cells in the peripheral blood of patients with gastric cancer.</p><p><b>METHODS</b>Peripheral blood was obtained from 40 patients with gastric cancer and from 20 healthy volunteers. The mRNA of MAGE-1 and MAGE-3 genes in the peripheral blood mononuclear cells (PBMC) was detected by RT-PCR. The expressions of MAGE-1 and MAGE-3 mRNA in the tumor tissues of these gastric cancer patients were also detected by RT-PCR. Meanwhile,CEA expression by nested RT-PCR in PBMC of 40 gastric cancer patients was also detected.</p><p><b>RESULTS</b>Of 40 gastric cancer patients, MAGE-1 and MAGE-3 mRNA were positive in 47.5% (19/40) and 25% (10/40) of PBMC respectively, and in 62.5% (25/40) and 30% (12/40) of gastric cancer tissues respectively. As a whole, in the PBMC of 40 gastric cancer patients, 25 (62.5%) samples were found to express at least one type of MAGE mRNA. In the patients whose tumors did not express MAGE-1 and/or MAGE-3 genes, the corresponding MAGE mRNA was also undetected in their PBMC. There was no expression of MAGE-1 or MAGE-3 gene in the PBMC from the 20 healthy donors. The positive rate of MAGE mRNA in PBMC was closely correlated with the tumor stage and lymph node metastasis (P <0.05). Positive rate of CEA gene expression was 32.5% (13/40) in the PBMC of 40 gastric cancer patients, 29 (72.5%)samples were detected to express at least one type of MAGE gene and CEA gene mRNA.</p><p><b>CONCLUSIONS</b>MAGE-1, MAGE-3 and CEA mRNA are specifically detected with high percentage in the PBMC of gastric cancer patients by RT-PCR. They could be used as specific tumor markers for the detection of the circulating gastric cancer cells, and the detection results may be helpful to evaluate the prognosis of gastric cancer patients.</p>

Expression of MAGE-B genes in hepatocellular carcinoma / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To investigate the expression of MAGE-B genes in hepatocellular carcinoma (HCC) in order to find new targets for immunotherapy.</p><p><b>METHODS</b>The expression of MAGE-B1, B2, A1 and A3 mRNA was detected using RT-PCR in HCC tissues and the corresponding adjacent non-HCC tissues from 47 HCC patients, 30 samples of cirrhosis and normal liver tissues. Four samples selected randomly from MAGE-B1 or B2 with positive RT-PCR results were sequenced to confirm the results of RT-PCR. The relationship between the expression of MAGE-B and some clinicopathological parameters was analyzed.</p><p><b>RESULTS</b>MAGE-B1 mRNA and MAGE-B2 mRNA were detected in 44.7% (21/47) and 61.7% (29/47) of HCC samples, respectively, while neither MAGE-B1 nor MAGE-B2 could be detected in the corresponding adjacent non-HCC liver tissues. In addition, none of 30 samples of cirrhosis and normal liver tissues was shown to express both MAGE-B genes. The DNA sequence confirmed that the RT-PCR products were truly target cDNA. The frequency of the expression of MAGE-A1 and A3 was 74.5% (35/47) and 44.7% (21/47), respectively. There was significant correlation between the expression of MAGE-B and MAGE-A (P < 0.05). However, the positive expression of MAGE-B was observed in 5 out of 12 HCC tissues without expression of MAGE-A1 and/or A3. When all four MAGE genes were examined, the positive rate of expression of one, two, three and four genes was 83.0% (39/47), 55.3% (26/47), 48.9% (23/47), and 38.3% (18/47) of 47 HCC tissues, respectively. No correlation was found between the expression of MAGE-B and clinical parameters such as age, sex, tumor size, degree of tumor differentiation, serum alpha-fetoprotein level and hepatitis B virus or hepatitis C virus infection (P > 0.05).</p><p><b>CONCLUSION</b>MAGE-B genes are expressed with relatively high frequency and specificity in HCC. Most HCC patients with positive expression of at least one member of MAGE-B or MAGE-A gene family are adequate candidates to receive specific immunotherapy. Frequent co-expression of multiple members of MAGE-B and MAGE-A subfamilies provides the possibility of using polyvalent vaccines to achieve more effective immunotherapeutic results.</p>

Expression and clinical significance of hepatocellular cancer antigen genes in human hepatocellular carcinoma / 中华外科杂志

<p><b>OBJECTIVE</b>To investigate the expression of four hepatocellular cancer antigen (HCA) gene mRNA in hepatocellular carcinoma.</p><p><b>METHODS</b>The expression of HCA90, HCA519, HCA520, HCA587 mRNA was detected using RT-PCR in HCC tissues and the corresponding adjacent non-HCC tissues from 46 HCC patients, cirrhosis tissues from 10 samples and normal liver tissues from 10 samples. The relationship between positive expression rate of HCA gene and clinical and lab data was evaluated.</p><p><b>RESULTS</b>Of 46 HCC tissues, HCA90, HCA519, HCA520 and HCA587 mRNA were detectable in 65.2%, 76.1%, 45.7% and 32.6%, respectively. At least one HCA gene mRNA was positive in 82.6% of HCC tissues. Only weak expression of HCA519 could be detectable in 6.5% of the corresponding adjacent non-HCC tissues. None of 10 samples of cirrhosis and normal liver tissues expressed any HCA gene mRNA. No correlation was found between the expression of HCA and clinical date such as age, sex, tumor size, tumor differentiation, serum alpha-fetoprotein level and hepatitis B virus infection or hepatitis C virus infection (P > 0.05). However, in some patients with normal serum alpha-fetoprotein (< 25 ng/L), specific expression of HCA genes was observed.</p><p><b>CONCLUSION</b>HCA gene mRNA is expressed with a high percentage and specificity in hepatocellular carcinomas and their products are new potential promising targets for immunotherapy of HCC.</p>

Expression of 5-hydroxytamine receptors in hepatic stellate cell and action of 5-hydroxytamine on biological characteristics of hepatic stellate cell / 中华外科杂志

<p><b>OBJECTIVE</b>To investigate the expression of 5-hydroxytamine receptors in hepatic stellate cells HSCs and action of 5-hydroxytamine on biological characteristics of HSC.</p><p><b>METHODS</b>Liver ex vivo perfusion of collagenase and density gradient centrifugation were used to isolate hepatic stellate cell. RT-PCR was used to detect the expression of 5-hydroxytamine receptor subtypes 1A, 2A, 2B and 3. Western blot hybridization was used to elucidate the effect of 5-hydroxytamine and its 2A receptor antagonist ketanserin and 3 receptor antagonist ondanosetron on expression of transforming growth factor-beta1 (TGF-beta1) and Smad4 in HSC. HSCs were cultured on silicone membrane. The effect of 5-hydroxytamine, ketanserin and ondanosetron on cell contraction were studied.</p><p><b>RESULTS</b>HSC expressed 5-hydroxytamine receptors subtypes 1A, 2A and 2B. 5-hydroxytamine significantly increased the expression of TGF-beta1 and Smad4 in HSC (P < 0.05). This was antagonized by ketanserin, not by ondanosetron. 5-hydroxytamine induced cell contraction in a dose-dependant manner. Ketanserin antagonized this action, but ondanosetron did not.</p><p><b>CONCLUSIONS</b>HSCs express 5-hydroxytamine receptors. 5-hydroxytamine could affect the biological characteristics of HSC through its receptor mediation, and may play a role in the pathogenesis of liver cirrhosis and portal hypertension.</p>