张春凤, 贾淑芹①, 苏秀兰①. 抗癌生物活性肽对BGC-823 细胞基因表达的影响*[J]. 中国肿瘤临床, 2010, 37(18): 1021-1023. DOI: 10.3969/j.issn.1000-8179.2010.18.001
引用本文: 张春凤, 贾淑芹①, 苏秀兰①. 抗癌生物活性肽对BGC-823 细胞基因表达的影响*[J]. 中国肿瘤临床, 2010, 37(18): 1021-1023. DOI: 10.3969/j.issn.1000-8179.2010.18.001
ZHANG Chunfeng1, JIA Shuqin2, SU Xiulan2. Effect of Anticancer Bioactive Peptide on the Gene Expression of Human Gastric Cancer BGC- 823 Cells[J]. CHINESE JOURNAL OF CLINICAL ONCOLOGY, 2010, 37(18): 1021-1023. DOI: 10.3969/j.issn.1000-8179.2010.18.001
Citation: ZHANG Chunfeng1, JIA Shuqin2, SU Xiulan2. Effect of Anticancer Bioactive Peptide on the Gene Expression of Human Gastric Cancer BGC- 823 Cells[J]. CHINESE JOURNAL OF CLINICAL ONCOLOGY, 2010, 37(18): 1021-1023. DOI: 10.3969/j.issn.1000-8179.2010.18.001

抗癌生物活性肽对BGC-823 细胞基因表达的影响*

Effect of Anticancer Bioactive Peptide on the Gene Expression of Human Gastric Cancer BGC- 823 Cells

  • 摘要: 目的:利用基因芯片技术分析一种新型抗癌生物制剂- 抗癌生物活性肽对人胃癌BGC-823 细胞的基因表达谱调控的影响,并对部分差异表达基因进行RT-PCR 检测,从而验证芯片的有效性。方法:订制表达谱芯片,分别将400 条和648 条人类基因的PCR 产物点样于特殊处理后的玻璃片上制备基因表达谱芯片。以加入不同浓度抗癌生物活性肽及处理不同时间的BGC-823 细胞为实验组,以生理盐水处理的BGC-823 细胞作为对照组,当确定最佳作用时间及抗癌生物活性肽有效作用浓度后,分别提取实验组与对照组细胞的总RNA,反转录成cDNA ,以两种荧光素Cy5 和Cy3 标记后作为探针,与制备好的表达谱芯片进行杂交,以生物信息学方法进行数据的处理和分析,并将其中的差异表达基因p15和HSP 701B 进行RT-PCR 的验证。结果:两张芯片上差异表达的基因共47个,其中上调27个,下调20个。这些基因按照功能分类涉及细胞周期、代谢酶、免疫相关、细胞凋亡、抑癌基因等类别,包括HSP 701B、HSP 75、HSP 90、磷脂酶D2(PLD 2)、凋亡抑制基因(IEX-1L)、人类杆状病毒IAP 重复序列3、E2 泛素耦联酶Ubch5c、人类缺氧诱导因子1 α 亚基、γ- 干扰素、TNF 诱导因子、人类黑色素瘤抗原p15基因等。同时RT-PCR 结果验证了芯片的检测的有效性。结论:抗癌生物活性肽具有调控人胃癌BGC-823 细胞基因表达的作用,其中对抑癌基因、热休克蛋白家族基因等有较明显的影响;基因芯片可以为阐明抗癌药物的作用机制提供有力的分子基础。

     

    Abstract: Objective:To analyze the influence of a new kind of anticancer biological preparation, anticancer bioactive peptide (ACBP), on gene expression profile alteration of the human gastric cancer cell line BGC- 823 using microarray tech -nique, clarify its anticancer mechanism and test its validity for part of the differentially expressed genes by using RT-PCR method. Methods:Two cDNA microarrays, containing 400 genes and 648 genes, were prepared by spotting purified PCR products onto specially treated glass slides. BGC-823 cells treated with different concentrations of ACBP at different times were used as the experimental group and BGC-823 cells treated with saline were used as the control group. After the best action time and concentration were determined, probes were made as follows: total RNA was extracted and reverse tran-scribed into cDNA for both the experimental and control groups, and labeled with Cy5 and Cy3 fluorescent dyes, respective -ly. These were then hybridized onto microarrays. Subsequently, data were analyzed using bioinformatics. RT-PCR was used to test candidate genes p 15and HSP701 B. Results:There were a total of47differentially expressed genes on the two microarrays: 27were up-regulated and20were down-regulated. These genes were classified into several categories: cell cycle, metabolic enzymes, immune related genes, cell apoptosis and tumor suppressor genes, including HSP701 B, HSP75, HSP 90, PLD 2, IEX- 1L, IAP3, Ubch5c, γ-IFN, p15. RT-PCR results showed that p 15 was up-regulated and HSP701 B was down-regulated in the experimental group compared with the control group, which also confirmed the validi -ty of the microarrays. Conclusion : Anticancer bioactive peptide has the function of regulating the gene expression of human gastric cancer cells,such as tumor suppressor genes and heat shock protein family members. Microarrays can provide a powerful molecular basis for clarifying the mechanism of anticancer drugs.

     

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