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Effect of YB-1 Downregulation Via RNAi on EGFR Expression and Cell Proliferation in Human Lung Adenocarcinoma Cell Line A549

Author: ZhaoYuDa
Tutor: GuChunDong
School: Dalian Medical University
Course: Thoracic Surgery
Keywords: Human lung adenocarcinoma cells YB-1 EGFR siRNA Cell proliferation
CLC: R734.2
Type: Master's thesis
Year: 2010
Downloads: 81
Quote: 1
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Abstract


Objective: In recent years, lung adenocarcinoma gradually replaced squamous cell lung cancer pathological type. Surgery and traditional put the efficacy of chemotherapy is reliable, but the benefits of its development into a relatively stable plateau, while targeted therapies and novel molecular targeted drug development success for the anti-tumor research bring good prospects. Human YB-1 protein (Y box-binding protein-1) is an important class of specific binding of the target gene promoter and enhancer internal Y-box sequences of nuclear transcription and translation factors. Epidermal growth factor receptor (EGFR) promoter region containing the sequence of the Y-box. Both have an important role in the pathology of the start of the tumor, proliferation, invasion and metastasis. On the above knowledge base, the experimental use of RNAi (RNA interference) technology, silent lung adenocarcinoma cell line A549 YB-1 protein expression, explore the YB-1 protein in silence before and after EGFR expression and cell proliferation differences, thereby determining YB-1 protein expression of EGFR whether the impact of regulation and its effect on cell proliferation. Methods: Lipofectamine2000 transfection reagent Santa Cruz production the YB-1siRNA fragment into A549 cells, the normal group a simple Lipofectamine2000 treated and Control siRNA group for control. In the selected YB-1 siRNA fragments exact effect on the basis of analysis by RT-PCR, Western Blot method of silent YB-1 groups before and after the YB-1 and EGFR gene and protein expression in A549 cells; YB-1 siRNA inhibition rate of cell proliferation by MTT assay. The above experimental data using SPSS11.5 statistical software. All experiments were repeated three times, Measurement data were expressed as mean ± standard deviation ((?) ± S) indicates whether the difference was significant (P lt; 0.05) using single-factor analysis of variance between the experimental group. Results: RT-PCR and Western Blot results show YB-1 mRNA and protein expression was significantly decreased in A549 cells after YB-1 siRNA interference EGFR mRNA and protein expression decreased (P lt; 0.05), preliminary confirmed lung adenocarcinoma cell line A549 YB-1 regulation of EGFR. YB-1 siRNA 2.MTT detection reaches a certain dose of interference on cell proliferation inhibition produced (0.0625μl, P gt; 0.05; 0.125μl, P lt; 0.05), and there is an optimal inhibitor amount. Conclusion: 1. Gene and protein by RT-PCR and Western Blot method preliminarily confirmed YB-1 protein expression of EGFR regulation. By MTT assay confirmed that YB-1 siRNA interference can inhibit tumor cell proliferation and there is an optimal dose.

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CLC: > Medicine, health > Oncology > Respiratory system tumors > Lung tumors
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