中国癌症研究（英文版）杂志

Objective: A single mechanistic pathway cannot explain the genesis of drug resistance in cancer. Drug resistance in cancer is a major obstacle to successful chemotherapy. KB cells provide a useful starting point for selection of the multidrug resistant (MDR) cell lines. Methods: We used cDNA microarrays containing 12,720 sequences of known genes, expressed sequence tags and unknown clones to monitor gene expression profiles in MDR KB cells. Results: Preliminary data analysis showed that 18 genes were up-regulated and 18 genes were down-regulated by comparison of expression patterns between KB 3-1 and MDR KB-V1 cells. Furthermore, the highly over-expressed CGA, CLU genes in MDR KB-V1 cell were verified with conventional Northern blot analysis. These genes contain information predictive of drug resistance of cancer cells. Conclusion: Our study demonstrates that genome-wide gene expression profiling by using cDNA microarray technique is a valuable approach in obtaining molecular mechanism of drug resistance in cancer cells.

作者：汪进；黄明辉；曾志雄；方宏勋；杨梦苏 刊期： 2002年第01期

Objective: To elucidate the expression and significance of cell cycle protein cyclin D1 in human hepatocellular carcinoma (HCC). Methods: The expression of cyclin D1 protein in 29 cases of HCC tissues was detected by immunohistochemical ABC method, and the relationship between its positive rate and pathological grades of HCC tissues was analyzed. Results: The positive rate of cyclin D1 in HCC tissues was 58.6%(17 in 29 cases), whereas only 18.2% (2 cases of 11 cases) in the non-tumor liver tissues immediately adjacent to HCC tissues (LAH). There was significant difference between grade II and LAH tissues (P<0.05), and between grade I and grade III on the positive rate of cyclin D1 (P<0.05), respectively. Conclusion: Cyclin D1 may be regarded as an oncogenic marker during the genesis and development of HCC, and its role in the transforming process from G1 phase to S phase of HCC cells needs further studies.

作者：杨连君；司晓辉 刊期： 2001年第02期

To investigate the significance of E- cadherin (E-cad) and CD44v6 expression in human hepatocellular carcinomas (HCCs). Methods: An immunohistochemical method was used to detect E-cad and CD44v6 expression in 66 cases of HCCs. Results: The positive rates of E-cad and CD44v6 expression in human HCCs were 42.4%(28/66) and 39.4%(26/66), respectively. There was an inverse correlation between E-cad expression and invasive and metastatic potential of HCCs (P<0.01), and a positive correlation between the CD44v6 expression and invasive and metastatic potential of HCCs (P<0.01). Moreover, the 5-year survival rate in the E-cad-positive group was higher than in E-cad-negative group (P<0.01), and that in the CD44v6-positive group was lower than in the CD44v6-negative expression group (P<0.05). Conclusion: these data show a possible association between E-cad and CD44v6 expression and the potential of invasion and metastasis in HCCs. E-cad and CD44v6 expression may be used as an auxiliary prognostic indicator in HCCs.

作者：郑建明；郑唯强；龚志锦；朱明华；戴益民；张照环 刊期： 2001年第04期

Tea polyphenols present in green tea show cancer chemopreventive effects in many tumor models. Epidemiological studies have also suggested that green tea consumption might be effective in the prevention of certain human cancers. In the present study, we investigated the molecular mechanisms of the inhibition of cell proliferation by tea polyphenols in nasopharyngeal carcinoma (NPC) cell line CNE1-LMP1. Methods: CNE1-LMP1 cells were treated with tea polyphenols at various doses (0, 25, 50, 100, 200 μg/ml) for 24 hours, the morphology of cells was observed by light microscopy, and cell survival rate was determined by MTT assay. At the same time, cell cycle of CNE1-LMP1 was analyzed by flow cytometry. Cyclin D1 transcription was analyzed by cyclin D1 promoterluciferase reporter system and expression of cyclin D1 protein by Western blot analysis. Transactivities of NF-kB and AP-1 was analyzed by Dual-fluorescence reporter gene system. Results: After treatment of CNE1-LMP1 cells with tea polyphenols, the number of proliferating cells was obviously decreased as determined by light microscopy and MTT assay (from 100% to 89.4%, 83.3%, 74.8% and 38.1%). With the increase of tea polyphenols concentrations, the number of cells in S-phase was obviously decreased, and the number of cells in G1-phase from 22.20% to 13.16%, and the number of cells in G0/G1 phase was elevated from 68.5% to 74.08%. It suggests that tea polyphenols could arrest the cell cycle at both of the two checkpoints. Furthermore, transcription and were obviously declined 7-8 folds (100-200 mg/ml tea polyphenols or EGCG group) and expression of cyclin D1 protein also decreased in a dose-dependent manner. Transactivities of NF-kB and AP-1 were obviously down-regulated in CNE1-LMP1 cells. Conclusion: Green tea polyphenols could inhibit cell proliferation, by suppressing the activity of NF-kB and AP-1, and by down-regulation of the transcription of cyclin D1.

作者：罗非君；胡智；邓锡云；赵燕；曾亮；董子刚；易薇；曹亚 刊期： 2001年第04期

Objective: To establish a fluoregenic probe quantitative RT-PCR (FQ-RT-PCR) method for detection of the expression of MDR1 gene in tumor cells and to investigate the expression of MDR1 gene in patients with lung cancer. Methods: The fluorogenic quantitative RT-PCR method for detection of the expression of MDR1 gene was established. K562/ADM and K562 cell lines or 45 tumor tissues from patients with lung cancer were examined on PE Applied Biosystems 7700 Sequence Detection machine. Results: the average levels of MDR1 gene expression in K562/ADM cells and K562 cells were (6.86±0.65)× 107copies/mg RNA and (8.49±0.67)×105 copies/mg RNA, respectively. The former was 80.8 times greater than the latter. Each sample was measured 10 times and the coefficient variation (CV) was 9.5% and 7.9%, respectively. Various levels of MDR1 gene expression were detected in 12 of 45 patients with lung cancer. Conclusion: Quantitative detection of MDR1 gene expression in tumor cells was achieved by using FQ-RT-PCR. FQ-RT-PCR is an accurate, and sensitive method and easy to perform. Using this method, low levels of MDR1 gene expression could be detected in 24% of the patients with lung cancer.

作者：高劲松；马刚；仝明；陈佩毅；王传华；何蕴韶 刊期： 2001年第02期

Objective: To compare the differential expression of mRNA between MKN-28 (highly differentiated) and MKN-45 (poorly differentiated) gastric adenocarcinoma cells and identify genes involved in human gastric adenocarcinoma differentiation. Methods: Differential expression of mRNA between MKN-28 and MKN-45 adenocarcinoma cells was investigated by fluorescent differential display (FDD). Differentially expressed cDNA was analyzed by bio-informatics and confirmed by RT-PCR and Northern-blot. Results: 45 differential fragments were finally attained. One of them (No. 10) was an approximate 750 bp cDNA and highly up-regulated in MKN-45 cells as compared with MKN-28 cells. By using Blastn and UniGene database analysis, we found the fragment was mapped to chromosome 14q11.2(q12 and showed a significant homology to Bcl-2 binding protein gene (BNip3), which was recently identified encoding pro-apoptosis protein located in mitochondrial. Conclusion: The BNip3 induced apoptosis could be suppressed by interacting with bcl-2. The BNip3 gene in tumor cells might be up-regulated by the hypoxia response element through the HIF1a transcription factor, causing death of the hypoxic cells at the center of the tumor where vascularization is usually poor in the process of tumor development.

作者：王建华；陈诗书 刊期： 2002年第01期

Objective: To investigate the effects of anti-PML (promyelocytic leukemia) or anti-PML/RARa (promyelocytic leukemia/retionic acid receptora) antisense oligonucleotides on cell growth, expression of PML-RARa mRNA and PML-RARa/PML protein location of NB4 cell lines. Methods: RT-PCR was used for detecting PML-RARa mRNA expression, trypan blue exclusion for cell count, methylcellose assay for leukemic colony forming unit detection, immuno- fluorescence for PML-RARa/PML protein location. Results: Both anti-PML start codon region antisence (STAS) and anti-PML-RARa fusion region antisence (FUAS) could inhibit cell growth and the formation of acute myelocytic colony forming unit of cells(AML-CFU). Cells become partial differentiated at days 5, being more obvious in FUAS-treated cells than in STAS ones. Down regulation of PML-RARa mRNA expression occurred at 24 hours in STAS and FUAS-treated cells and maintained for up to 72 hours. Immuno-fluorescence analysis with anti-PML monoclonal antibody showed a remarkable decrease even complete disappearance of microgranules. The residual granules became enlarged as discrete dots (<10 per cell), similar to normal POD structure in some STAS-treated cells at 24 hours. At 72 hours, nearly all the granules disappeared. Similar changes were observed in FUAS-treated cells. Conclusion: Both PML and PML-RARa antisence oligonucleotides can specially block the expression of PML-RARa at mRNA and protein levels. PML protein is implicated in the regulations of cell differentiation.

作者：陈烨；缪金明；方智雯；朱学宏；邵念贤；欧阳仁荣 刊期： 2001年第02期

To explore the death-related factors of stageⅠrectal cancer patients. Methods: 89 cases of stage I rectal cancer patients between 1985 and 2000 were retrospectively studied for prognostic factors. Factors including age, gender, tumor size, circumferential occupation, gross type, pathological type, depth of tumor invasion, surgical procedure, adjuvant chemotherapy and postoperative complication were chosen for cox multivariate analysis (forward procedure) using Spss software (10.0 version). Results: multivariate analysis demonstrated that muscular invasion was an independent negative prognostic factor for stageⅠrectal cancer patients (P=0.003). Conclusion: Muscular invasion is a negative prognostic factor for stage I rectal cancer patients.

作者：武爱文；顾晋；薛钟麒；王怡；徐光炜 刊期： 2001年第04期

Objective: To introduce the method of a modified transcranial approach for resection of paranasal sinuses tumors involving the anterior skull base and to address our experience with the approach. Patients and Methods: Ten cases were operated by the approach. Among them, 4 suffered from benign meningeomas, 6 with malignant tumors included one chondrosarcoma, two malignant meningeomas, two olfactory neuroblastomas, and one squamous sarcoma. Of the patients, 4 cases had primary tumor and 6 cases had recurrent tumors. Result: All of the ten cases underwent operation and no postopertion complication occurred. 7 cases have survived for one to four years without tumor recurrence. 3 cases with malignant tumor died of tumor relapse in one to two years. Conclusion: This method significantly has helped to reduce the persistence and recurrence of the disease.

作者：赵素萍；陶正德；肖健云 刊期： 2001年第02期

Objective: To study the effect of electromagnetic pulse (EMP) on apoptosis of human lung carcinoma cell line GLC-82. Methods: The injury changes in GLC-82 cells after irradiated by EMP (electric field intensity was 60 kV/m with 5 pulses/2 min) were analyzed by cytometry, MTT chronometry and flow cytometry. The immuno- histochemical SP staining was used to determine the expressions of bcl-2 protein and p53 protein. The stained positive cells were analyzed by CMIAS-II image analysis system at a magnification 400. All data were analyzed by SPSS8.0 software. Results: EMP could obviously inhibited lung carcinoma cell line GLC-82 proliferation and increased the number of non-adherent cells. The absorbance value (A570) of MTT decreased immediately, at 0 h, 1 h and 6 h after the GLC-82 cells irradiated by EMP as compared with control group. The highest apoptosis rate was found to reach 13.38% by flow cytometry at 6 h after EMP irradiation. Down-regulation of bcl-2 expression and up-regulation of bax expression were induced by EMP. Conclusion: EMP promoted apoptosis of GLC-82 cells. At same time, EMP can down-regulate bcl-2 expression and up-regulate p53 expression in GLC-82 cells. The bcl-2 and the p53 protein may involve the apoptotic process.

作者：曹晓哲；赵梅兰；王德文；董波 刊期： 2002年第01期