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Differential expression of a novel colorectal cancer differentiation-related gene in colorectal cancer

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World Journal of Gastroenterology 2001年第4期7卷 551-554页

作者： Xing-Guo Li Jin-Dan Song Yun-Qing Wang Key Laboratory of Cell Biology,Ministry of Public Health of China,China Medical University,Shenyang 110001,Liaoning Province,China Key Laboratory of Cell Biology Ministry of Health of China China Medical University Shenyang 110001 Liaoning Province China.

AIM: To investigate SBA2 expression in CRC cell lines and surgical specimens of CRC and autologous healthy mucosa. METHODS: Reverse transcription-polymerase chain reaction (RT-PCR) was used for relative quantification of SBA2 mRNA levels in 4 human CRC cell lines with different grades of differentiation and 30 clinical samples. Normalization of the results was achieved by simultaneous amplification of beta-actin as an internal control. RESULTS: In the exponential range of amplification, fairly good linearity demonstrated identical amplification efficiency for SBA2 and beta-actin (82%). Markedly lower levels of SBA2 mRNA were detectable in tumors, as compared with the coupled normal counterparts PP P<0.05). CONCLUSION: SBA2 gene might be a promising novel biomarker of cell differentiation in colorectal cancer and its biological features need further studies.

关键词： Gene Expression Regulation, Neoplastic Cell Differentiation Colorectal neoplasms DNA, Complementary Humans Molecular Sequence Data neoplasm proteins Polymerase Chain Reaction Tumor Cells, Cultured Tumor Markers, Biological

Induction of cytotoxic T lymphocytes from the peripheral blood of a hepatocellular carcinoma patient using melanoma antigen-1 (MAGE-1) peptide

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Chinese Medical Journal 2002年第7期115卷 1002-1005,145-146页

作者：吕建锋冷希圣彭吉润牟东成庞学雯商小英陈慰峰北京大学人民医院肝胆外科北京100044 北京大学医学部免疫学系北京100044

OBJECTIVE: To investigate the possibility of using melanoma antigen-1 (MAGE-1) peptide as a tumor vaccine to treat hepatocellular carcinoma (HCC). METHODS: The expressions of MAGE-1 in 8 HCC cell lines and in liver cancer tissue from a patient were detected using RT-PCR. The type of human leucocyte antigen I(HLA I) of both 8 HCC cell lines and peripheral blood mononuclear cells of the patient was detected using a microcytotoxicity method to screen out target cell lines for the cytotoxicity assay. Peripheral blood mononuclear cells from the HCC patient pulsed with an MAGE-1 peptide (NYKCRFPEI) were used as antigen presenting cells. Autogenous peripheral blood mononuclear cells were stimulated with antigen presenting cells every 7 days for 4 times to elicit cytotoxic T lymphocytes. The phenotype of effector cells was analyzed using flow cytometry. The cytotoxicity of effector cells was detected with a lactate dehydrogenase releasing assay. RESULTS: The expressions of both MAGE-1 and HLA-A24 were detected in BEL7405 cell line which were used as the positive target cell line in the cytotoxicity assay. The expression of MAGE-1 alone was detected in HLE, BEL7402, BEL7404, QGY7703 and SMMC7721 cell lines, and the expression of neither MAGE-1 nor HLA-A24 was shown in QGY 7701 and HpG2 cell lines. The last 7 cell lines could be used as negative target cell lines in the cytotoxicity assay. Peripheral blood mononuclear cells expanded 32 folds during 28-day culture. The ratio of CD3(+) T cells increased by 16% (from 54% to 70%), and the ratio of CD8(+) T cells increased by 20% (from 36% to 56%) during 28-day culture. When the ratio of effector cells to target cells was 10:1, effector cells exhibited 62.5% cytotoxicity against autogenous lymphoblasts pulsed with the peptide (NYKCRFPEI) of MAGE-1 antigen, 40.25% cytotoxicity against BEL7405 cells, compared with 17.88% cytolysis observed against autogenous lymphoblasts, 19.55% against HLE cells, and 1.6% against QGY7701 cells. Whe

关键词： Adult Cancer Vaccines Carcinoma, Hepatocellular HLA-A Antigens Humans Liver neoplasms Male neoplasm proteins RNA, Messenger T-Lymphocytes, Cytotoxic Tumor Cells, Cultured

Mcl-1 mediates cytokine deprivation induced apoptosis of human myeloma cell line XG-7

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Chinese Medical Journal 2002年第8期115卷 1241-1243页

作者：宋伦黎燕孙英勋沈倍奋军事医学科学院基础医学研究所北京100850

OBJECTIVE: To investigate apoptosis in XG-7, a human myeloma cell line, induced by IL-6 deprivation and the function of three anti-apoptotic Bcl-2 proteins (Bcl-2, Bcl-kappa(L), Mcl-1) in the apoptotic process. METHODS: Apoptosis in XG-7 myeloma cells induced by IL-6 withdrawal was determined by flow cytometry with propidium iodide (PI) nuclear staining. Expressions of three Bcl-2 proteins in XG-7 cells were monitored by immunoblotting assay. RESULTS: In the absence of IL-6 for a certain time, a significant percentage of apoptiotic XG-7 cells can be observed, as well as down-regulated expression of one of the three anti-apoptotic proteins (Mcl-1) in XG-7 cells. IL-6 re-stimulation in XG-7 cells following cytokine removal up-regulated the expression of Mcl-1 and inhibited cell apoptosis. CONCLUSION: Mcl-1,instead of Bcl-2 and Bcl-kappa(L), plays an important role in IL-6 deprivation induced apoptosis in XG-7 human myeloma cells.

关键词： Apoptosis Humans Interleukin-6 Multiple Myeloma neoplasm proteins Proto-Oncogene proteins c-bcl-2 Research Support, Non-U.S. Gov't Tumor Cells, Cultured bcl-X Protein

Increased sensitivity of colorectal cancer cell lines with microsatellite instability to 5-fluorouracil in vitro

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Chinese Medical Journal 2002年第7期115卷 1048-1052,151-152页

作者：陈修煦来茂德黄琼浙江大学医学院病理学教研室杭州310031

OBJECTIVE: To study the relationship between sensitivity to 5-FU and the status of a panel of microsatellite loci in three human colon cancer cell lines. METHODS: Cell viability in several concentrations of 5-FU was assessed by the MTT test. Expression of hMSH2 and hMLH1 in LoVo, SW480 and SW1116 cells were analyzed by immunocytochemical *** mononucleotide and dinucleotide microsatellite loci were analyzed by the PCR-SSLP-silver staining method. RESULTS: By MTT assay, it showed that LoVo cells were more sensitive than SW480 and SW1116 cells (0.8 micromol/L,2.2 micromol/L and 1.9 micromol/L, respectively, P

关键词： DNA-Binding proteins Microsatellite Repeats Antimetabolites, Antineoplastic Base Pair Mismatch Carrier proteins Colorectal neoplasms DNA Repair Fluorouracil Humans Immunohistochemistry MutS Homolog 2 Protein neoplasm proteins Nuclear proteins Polymerase Chain Reaction Proto-Oncogene proteins Tumor Cells, Cultured