| Diffuse large B-cell lymphoma(diffuse large B-cell lymphoma, DLBCL) is a group of different clinical and biological characteristics of heterogeneous tumors, accounting for non-Hodgkin’s lymphoma(non-hodgkin’s lymphoma, NHL) 40 %, which up to 60% in developing countries, while conventional treatment methods can only 40-50% of patients continued to obtain clinical remission. Therefore, the search for new molecular markers in clinical has a positive meaning to their diagnosis and treatment.micro RNA as a single packet of small non-coding strand of RNA, a length of about 22-25 base pairs, After transcripts of genes these RNA play a regulatory role, according to inhibit or degradation protein it be divided into two functions. Studies have shown that, mi RNAs plays an vital role in the immune system regulation, the proliferation and differentiation and growth of cells. Its also plays a certain role in promoting or inhibiting of tumor development. In different tumors, different mi RNAs have different expression, so it can be used the biomarkers in clinical diagnosis, prognosis and treatment in clinicly. Studies have shown that mi R-29 b as a member of micro RNA, it can silence some potential oncogenes, inhibition of tumor cell growth, induction of apoptosis. The study also showed that, mi R-29 b functionality exists significant heterogeneity, mi R-29 b lack of corresponding resrarch in DLBCL expression and function. The study analyzed the expression and function of mi R-29 b in DLBCL cell lines, and investigated the role and mechanism of mi R-29 b in DLBCL development. ObjectLearning DLBCL cell lines for the expression of mi R-29 b.Observation of mi R-29 b mimic and mi R-29 b inhibitor in two DLBCL cell lines SU-DHL-8, the transfection rate case SU-DHL-10.To investigate the impact of mi R-29 b on the proliferation of both cell lines SU-DHL-8, SU-DHL-10.Study the migration which mi R-29 b on both SU-DHL-8 and SU-DHL-10 cell.Researching the affect of mi R-29 b on both SU-DHL-8 and SU-DHL-10 cell. Methods1、 Trizol reagent extraction utilizing normal human lymphocytes and lymph node tissue total RNA, mi R-29 b in accordance with the design of the reverse transcription primer and q RT-PCR primers; q RT-PCR method to detect the use of DLBCL cell lines for the expression of mi R-29 b.2、 according to mimic and inhibitor mi R-29 bsequence synthesized mi R-29 b is 3, the synthetic mi R-29 b of the mimic and inhibitor were transfected into cell lines by quantitative PCR for expression of mi R-29 b measured.3、mi R-29 b detect the impact on DLBCL cell line proliferation by CCK8 method.4、using the impact detection mi R-29 b on Transwell migration DLBCL cell lines, and the detection of cell apoptosis by flow cytometry situation. Results1. the results of mi R-29 b expression by quantitative PCR detection of DLBCL cell lines Display, mi R-29 b in human normal lymph node tissue was significantly higher than SU-DHL-10 and SU-DHL-8 cell lines expression(P <0.05).2. mi R-29 b mimic and inhibitor showed transfection efficiency, different concentrations of inhibitor,In the number of cells containing fluorescent cells is not very different(P> 0.05); and for confused with the increase, the number of cells transfected with mi R-29 b mimic a significant increase(P <0.05).3. Were transfected with mi R-29 b mimic and mi R-29 b inhibitor can alter the expression of mi R-29 b of DLBCL cells,and the expression level of mi R-29 b as compared to normal, mimic expression level of mi R-29 b improves 200-300 times,there are significant differences in the results(P <0.05),inhibitor and the control group compared to inhibit the expression of mi R-29 b make mi R-29 b reduce the expression of about 50%(P <0.05).4.CCK-8 cell growth assay mi R-29 b on SU-DHL-8, SU-DHL-10 cell line showed proliferation, the results showed that: compared with control group, increased mi R-29 b could significantly inhibit cell proliferation(P <0.05), and reduced mi R-29 b makes the proliferation of cells was significantly enhanced(P <0.05).5. With effect Transwell migration assay mi R-29 b on the migration of DLBCL cell lines. The results showed that: Compared with control group, increased mi R-29 b could significantly inhibit cell migration(P <0.05), and reduced mi R-29 b is the migration of cells was significantly enhanced(P <0.05). Conclusions1. Down-regulated expression of mi R-29 b was speculated that it might be involved in the development of diffuse large B cell lymphoma.2. Mi R-29 b can play a role of tumor suppressor gene in DLBCL, which is expected to become a new mi RNA-based tumor intervention agent, which needs further study. |
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