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The Role and Possible Mechanisms of miR-125b in Drug Resistance of Gastric Cancer Cell Lines

Author: ZhiHui
Tutor: LiuPing
School: Nanjing Medical University
Course: Oncology
Keywords: miR-125b Gastric Cancer Resistance BCL2 MCL1
CLC: R735.2
Type: Master's thesis
Year: 2011
Downloads: 155
Quote: 0
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Abstract


In recent years, clinical tumor drug resistance has become an important issue facing cancer treatment, one study tumor cell drug resistance mechanisms and to explore ways of reversing drug resistance is an important field of cancer research direction. Tumor drug resistance can be divided into primary and acquired drug resistance, a common clinical mostly acquired resistance. Now that the primary drug resistance and tumor genetics is closely related to the level of variation, while tumors with acquired resistance level of epigenetic gene variant are closely related. Epigenetics does not involve DNA sequence changes, mainly through DNA methylation sites, histone modifications and non-coding RNA regulation three aspects related gene expression regulation in tumor generation mechanism of acquired resistance may play an important role. microRNA (miRNA) as a newly discovered post-transcriptional regulation of important molecular extensively involved in various biological processes, such as: biological evolution and development, cell differentiation and malignant transformation, cell proliferation and apoptosis. In recent years, miRNA and cancer drug research rapidly changing relationship to tumor resistance mechanism provides new ideas, increase or inhibit tumor resistance associated with regulation of miRNA expression in tumor resistance function. MiRNA regulation currently reported less resistant to gastric cancer, studies have found that: miR-15b, miR-16 and miR-181b, miR-497 in gastric cancer multidrug resistant strains SGC-7901/VCR low expression, respectively. targeted inhibition of these miRNAs may increase the anti-apoptotic protein BCL2 thus partially reverse cell multidrug resistance phenotype. For further study microRNA-resistant cells in gastric cancer and molecular mechanisms of action, the experiment carried out the following studies. Method 1. Using miRNA miRNA microarray expression profiling combined real-time fluorescence quantitative RT-PCR detection of drug-resistant cancer cells were compared SGC7901/VCR and their parental cells SGC7901 expression differences between miR-125b. (2) the use of transient transfection of miR-125b mimics the way interventions resistant cells in the expression of miR-125b by MTT assay in vitro drug sensitivity assay on the cell resistant phenotype. 3 using the luciferase gene target validation experiments clearly miR-125b target genes and by Western blot and flow cytometry analysis miR-125b control the internal mechanism of resistance. Results 1. SGC7901/VCR cells miR-125b expression level was significantly lower than the parental cells, anti-apoptotic protein BCL2 and MCL1 expression levels were significantly higher than the parental cells. 2 transfected miR-125b mimics resistant cells can be significantly increased expression levels of miR-125b, and a significant increase in drug-resistant cells to chemotherapeutic drug sensitivity. 3 luciferase experiments confirmed that BCL2, MCL1 is miR-125b target genes. 4 transfected with miR-125b mimics resistant cells BCL2, MCL1 protein expression levels were significantly lower than the control was transfected resistant cells and transfected with miR-125b mimics resistant cells to chemotherapy-induced apoptosis The sensitivity was significantly higher than control was transfected resistant cells. Conclusion miR-125b in gastric cancer cells resistant to vincristine expression level was significantly lower than the corresponding parental cells, miR-125b is downregulated by anti-apoptotic protein BCL2, MCL1 mediated gastric cancer cells to a variety of chemotherapy drug resistance.

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CLC: > Medicine, health > Oncology > Gastrointestinal Cancer > Gastric neoplasms
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