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Analysis of Radiosensitivity Relative Genes for Cervical Cancer Cell Lines

Author: YanYuFeng
Tutor: ShengXiuGui
School: Jinan University
Course: Oncology
Keywords: Cervical Cancer Cell lines Gene chip Radiosensitivity Differentially expressed genes RT-PCR
CLC: R737.33
Type: Master's thesis
Year: 2011
Downloads: 73
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Abstract


【Objective】 Cervical cancer is one of the most common female malignancy worldwide, and its incidence ranks second (after breast cancer), 78% of cases in the developing world, the cause of death for our female tumors second place; treatment of cervical cancer, in principle, mainly to surgery and radiation therapy, assisted with chemotherapy; in the late (Ⅱ b-Ⅳ) is more of concurrent chemoradiotherapy-based, but the 5-year survival rate is about 50% . Although radiotherapy is the main treatment for cervical cancer, but there are still a considerable number of patients relapse during the year, about 35% of invasive cervical cancer after treatment relapse; these patients can improve the effect of radiation therapy, it is necessary to study cervical radiosensitivity of cancer. Studies have shown that cancer patients for radiotherapy reaction depends on the individual's inherent radiosensitivity of tumor inherent radiosensitivity and radiation-induced gene expression and gene regulation related, find cervical cancer radiosensitive closely related gene develop individualized treatment plan for patients with cervical cancer, the better you can reduce the recurrence. In this study, training three different types of pathology differentiation of human cervical cancer cells, a single X-ray irradiation using a different agent, the analysis of the size of its survival preliminary judgment radiosensitivity and sensitivity; application of the human genome sequence oligonucleotide chip to obtain two different radiosensitivity of cervical cancer cells gene changes, through changes of gene expression profiles in cervical carcinoma cell lines before and after irradiation, the analysis is differentially expressed genes, and after a preliminary verification, hope to find radiosensitivity of cervical cancer-related genes. 【Methods】 train three human cervical cancer cell line Hela (adenocarcinoma), Siha (poorly differentiated squamous cell carcinoma) and Hcc94 (well-differentiated squamous cell carcinoma), given in three different doses of cells single 6MV X-ray irradiation, dose order 0,2,4,6,8,10 Gy, visible changes in the morphology of cervical cancer cell lines before and after irradiation; calculating cell colony formation rate, using SigmaPlot software \survival curve, can be obtained the radiosensitivity indicator: 2Gy after irradiation cell survival fraction (SF2), the parameters extrapolated values ??(N), the mean lethal dose (Do), and the like; before the use of flow cytometry for cell X-ray irradiation and the cycle and the proportion of apoptotic cells after irradiation, filter out the the radiosensitive differences cell lines. CapitalBio Corporation of human genome-wide expression of the gene expression profile of the cell line the sensitive difference before and after the the microarray detection 6Gy X-ray irradiation, differentially expressed genes in the screening before and after irradiation. By analyzing the genetic changes of the cell lines before and after X-ray irradiation to differences in expression of genes induced by X-ray. Four randomly selected differentially expressed genes of more than 2 times, use the relative real-time quantitative the fluorescence PCR validation chip genetically modified, to determine chip detection result is reliable. [Results] after irradiation of sub cervical cancer some cells were apoptotic changes appear rounded and shedding; irradiation after cell culture continued to train about 14 2 1d, the naked eye can see cell clones. SF2 value of three cervical cancer cell survival curve is obtained by fitting the dose Hela cell line 0.725 is the highest, Siha cell line 0.581 hcc94 cells to 0.391, belonging to the lowest, the difference was statistically significance (P lt; 0.05) . Flow cytometric analysis of the G2 / M phase fraction of the Hela cells before irradiation significantly lower than the 10.42% Hcc94 cell ratio 18.77% (P lt; 0.05); after 6Gy X-ray irradiation, G2 / M phase were significantly block, S The percentage of cells also declined. 24 hours after X-ray irradiation, Hela cells G 2 / M phase was significantly in than Hcc94 cells (42.9% VS30.95%, P lt; 0.05); 2 cells before irradiation The apoptosis rate increase. The results of the analysis of gene chip visible Hela cells after irradiation and before irradiation total of 314 differentially expressed genes, including genes upregulated 17, down 297; Hcc94 cell lines according to differences in genes for 111; 2-fold increase in gene 72 months, 39 down-regulated genes. Screened by analyzing 55 differentially expressed genes in more than double, to understand its molecular function. Random selection of more than four times the detection of differentially expressed genes relative RT-PCR technique, the amount of expression product, the results of RT-PCR results consistent with chip results prove chip results are reliable. 【Conclusion】 cervical carcinoma Hela cell radiosensitivity lowest, belonging to the radiosensitive cells, Hcc94 the highest radiosensitivity, select two cells in human whole genome expression microarray detection, by analyzing the radiotherapy-induced differences genes differentially expressed genes involved in DNA damage and repair, apoptosis and signal transduction and periodic adjustment of different molecular functional classification, some differences in gene even participate in more than one process. Closely related to radiosensitivity of cervical cancer may be related to the difference of the final screening expressed genes is significantly up-regulated genes in the radiation resistance of Hela cells, help us to identify cervical cancer radiation sensitivity or tolerance gene for radiation therapy improved sensitivity and provide new targets and methods.

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