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The Expression Adn Fuction of CHD5 Gene of Cervical Cancer

Author: ChenNi
Tutor: LaiGuoQi
School: Chongqing Medical University
Course: Zoology
Keywords: CHD5 RNA interference Cervical Cancer HeLa
CLC: R737.33
Type: Master's thesis
Year: 2011
Downloads: 22
Quote: 0
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Abstract


Objective: CHD5 (chromodomain helicase DNA-binding 5) in a variety of tumor gene deletion, mutation or methylation, including cervical cancer, the genes important role in the inhibition of cancer development. The study showed that the CHD5 gene by p19Arf/p53 p16INK4a/pRb ways to regulate cell proliferation, senescence and apoptosis. Order to further understand the relationship of the the CHD5 gene and cervical cancer by immunohistochemical methods to detect the expression of the the CHD5 gene in cervical tissue, for using CHD5/RNAi the intervention of cervical cancer cells HeLa MTT method and flow cytometry CHD5 gene expression down the distribution of cell proliferation and cell cycle of HeLa cells to explore the CHD5 with cervical cancer relationship. Method: 1. CHD5 protein expression using immunohistochemistry chemical method of detection of cervical cancer and normal cervical tissue; 2. Constructed a recombinant adenoviral vector for the CHD5 gene design siRNA sequences. Then infected HeLa cells by RT-PCR and western blot detection of interference effects; 3.MTT method to detect changes in cell growth and proliferation in recombinant adenovirus infected HeLa cells, flow cytometry recombinant adenovirus in HeLa cell cycle distribution impact. Results: 1. Immunohistochemistry CHD5 protein expression in normal tissue of the cervix and cervical cancer paraneoplastic organization was significantly higher than that of cervical cancer tissue (P lt; 0.05). CHD5 protein expression and clinicopathological features of the data found no correlation. Successfully constructed the CHD5 gene RNA interference recombinant adenovirus, adenovirus titer of 5 × 10 11 pfu / ml, infected HeLa cells can reduce CHD5 expression. 3.MTT detection CHD5 gene interference can promote the proliferation of HeLa cells (P lt; 0.05). Flow cytometry results showed the infection CHD5siRNA recombinant adenovirus, HeLa cell cycle changes, the decrease in G0/G1 phase cells (P lt; 0.05), an increase in S-phase cells (P lt; 0.05), the G2 / M phase cells with the control group was no significant difference (P gt; 0.05). Conclusion: 1.CHD5 protein cervical cancer expression down prompted the CHD5 the expression level of cervical cancer is closely related. 2.CHD5siRNA recombinant adenovirus successfully constructed. 3.CHD5siRNA recombinant adenovirus infected HeLa cells results suggest that the occurrence of cervical cancer may CHD5 expression down easier HeLa cells caused by the G1 / S checkpoint into S phase, causing excessive cell proliferation related.

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CLC: > Medicine, health > Oncology > Genitourinary tumors > Female genital tumors > Uterine tumors
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