本文選題：即刻早期反應(yīng)基因-1　切入點(diǎn)：輻射誘導(dǎo)性　出處：《鄭州大學(xué)》2014年碩士論文　論文類型：學(xué)位論文

【摘要】：背景和目的 子宮頸癌是婦科三大常見(jiàn)的惡性腫瘤之一,嚴(yán)重威脅著女性的健康。隨著對(duì)腫瘤的發(fā)生、發(fā)展機(jī)制的不斷深入，基因治療成為繼手術(shù)、放療、化療等傳統(tǒng)治療方式后的新的治療手段。腫瘤基因-放射治療也成為腫瘤治療研究的新熱點(diǎn)。 即刻早期反應(yīng)基因-1(Immediate Early Response Gene X-1，IEX-1),屬于即刻早期基因（IEG）家族之一，定位于人6號(hào)染色體短臂(6p21.3)，是一種能在生長(zhǎng)因子、紫外線、電離輻射及缺血缺氧等誘導(dǎo)下快速表達(dá),調(diào)控細(xì)胞的生長(zhǎng)、增殖、分化和凋亡等各方面的基因。IEX-1的在不同組織中的過(guò)表達(dá)可正性或負(fù)性影響細(xì)胞的生存和細(xì)胞進(jìn)程。有研究表明IEX-1mRNA的表達(dá)在受到紫外線照射之后顯著增加，具有時(shí)間和劑量依賴性。IEX-1的UVB誘導(dǎo)性使得其成為腫瘤基因治療的靶點(diǎn)具有可能性,也為其帶來(lái)了潛在的臨床應(yīng)用性。 研究證實(shí)IEX-1在多種腫瘤組織中低表達(dá)或表達(dá)消失；但其在宮頸癌組織中的表達(dá)情況尚罕有報(bào)道。本研究采用紫外線照射方法照射宮頸癌HeLa細(xì)胞，分析宮頸癌細(xì)胞中IEX-1基因的輻射可誘導(dǎo)性，同時(shí)應(yīng)用免疫組化的方法從蛋白水平分別檢測(cè)IEX-1、HIF-1α、NF-κB在宮頸癌組織,宮頸CIN組織及正常宮頸組織的表達(dá)情況及三者間的關(guān)系，進(jìn)一步探討IEX-1與宮頸癌發(fā)生發(fā)展的聯(lián)系，為其作為臨床治療宮頸癌的靶點(diǎn)基因提供依據(jù)。 方法 1．采用不同劑量紫外線照射人宮頸癌HeLa細(xì)胞，RT-PCR檢測(cè)照射后細(xì)胞IEX-1mRNA的表達(dá)情況，選出最佳紫外線劑量。 2．以最佳紫外線照射劑量照射人宮頸癌HeLa細(xì)胞，RT-PCR檢測(cè)照射后不同時(shí)間點(diǎn)細(xì)胞中IEX-1mRNA的表達(dá)情況。 3．應(yīng)用免疫組織化學(xué)SP法檢測(cè)50例宮頸癌患者、40例CIN及26例正常宮頸組織中IEX-1、HIF-1α、NF-κB蛋白的表達(dá)情況，并分析三者之間的關(guān)系。 4．應(yīng)用SPSS16.0進(jìn)行統(tǒng)計(jì)學(xué)分析，χ2檢驗(yàn)和Spearman等級(jí)相關(guān)分析,檢驗(yàn)水準(zhǔn)α=0.05,以P＜0.05為差異有統(tǒng)計(jì)學(xué)意義。 結(jié)果 1．分別應(yīng)用不同等劑量的UVB照射人宮頸癌HeLa細(xì)胞后，，IEX-1mRNA表達(dá)量均有明顯增高（P0.05）。以40mJ/cm2的UVB劑量照射細(xì)胞后IEX-1mRNA表達(dá)增高幅度最大，為最佳紫外線照射劑量。 2．以最佳紫外線劑量40mJ/cm2照射人宮頸癌HeLa細(xì)胞，培養(yǎng)不同時(shí)間后檢測(cè)IEX-1mRNA表達(dá)的表達(dá)量，培養(yǎng)4小時(shí)后IEX-1mRNA表達(dá)量最高（P0.05）。 3．IEX-1在宮頸癌組織中的陽(yáng)性表達(dá)率明顯低于宮頸CIN和正常宮頸組織,差異具有統(tǒng)計(jì)學(xué)意義(P0.05)；HIF-1α和NF-κB在宮頸癌組織中的陽(yáng)性表達(dá)率均明顯高于宮頸CIN和正常宮頸組織,差異具有統(tǒng)計(jì)學(xué)意義(P0.05)。 4．宮頸癌組織中IEX-1、HIF-1α、 NF-κB的表達(dá)均與組織學(xué)分級(jí)及有無(wú)淋巴結(jié)轉(zhuǎn)移相關(guān)(P0.05),與患者年齡、腫瘤的組織學(xué)類型及FIGO分期無(wú)關(guān)(P0.05)。 5．宮頸癌組織中HIF-1α和NF-κB的表達(dá)呈正相關(guān)（r=0.398,P0.05）,兩者同IEX-1的表達(dá)呈負(fù)相關(guān)(r1=-0.322,P0.05；r2=-0.517,P0.05)。 結(jié)論 1．人宮頸癌HeLa細(xì)胞中IEX-1具有輻射誘導(dǎo)性，可被UVB誘導(dǎo)表達(dá)，且具有時(shí)間和劑量依賴性。 2．IEX-1在宮頸癌組織中低表達(dá)，與NF-κB、HIF-1α表達(dá)呈負(fù)相關(guān)，三者共同調(diào)節(jié)腫瘤細(xì)胞的生長(zhǎng)和凋亡。
[Abstract]:Background and purpose
Cervical cancer is one of the three most common gynecologic malignant tumor, a serious threat to women's health. With the tumor occurrence, development mechanism deeply, gene therapy has become following surgery, radiotherapy, chemotherapy and other new treatment after conventional treatment. Tumor gene radiotherapy has become a new hotspot in cancer treatment.
Immediate early response gene -1 (Immediate Early Response Gene X-1, IEX-1), which belongs to the family of immediate early genes (IEG), located on the short arm of chromosome 6 (6p21.3), is a growth factor in the ultraviolet, ionizing radiation, hypoxia and ischemia induced the rapid expression, regulation of cell growth the proliferation in different tissues. The overexpression of positive or negative effects of cell survival and cell process of.IEX-1 gene in all aspects of differentiation and apoptosis. The study showed that the expression of IEX-1mRNA increased significantly after exposure to ultraviolet light, a time and dose dependent induction of.IEX-1 UVB has become a target for tumor gene therapy has the possibility, also brings the potential for its clinical application.
Study confirmed that low expression of IEX-1 in tumor tissue or expression disappeared; but its expression in cervical cancer tissues of Shanghan have been reported. This study used the irradiation of cervical cancer HeLa cells induced by ultraviolet irradiation method, analysis of radiation IEX-1 expression in cervical cancer cells, and immunohistochemistry at protein level were detected IEX-1, HIF-1 alpha, NF- kappa B in cervical cancer, the relationship between the expression of CIN in cervical tissue and normal cervical tissue and between the three, to further explore the occurrence and development of IEX-1 and cervical cancer, and provide the basis for the clinical treatment of cervical cancer as a target gene.
Method
1. the HeLa cells of human cervical cancer were irradiated with different doses of ultraviolet radiation. The expression of IEX-1mRNA in the cells after irradiation was detected by RT-PCR, and the best ultraviolet dose was selected.
2. the human cervical cancer HeLa cells were irradiated with the best ultraviolet radiation dose, and the expression of IEX-1mRNA in the cells at different time points after irradiation was detected by RT-PCR.
3. immunohistochemical SP method was used to detect the expression of IEX-1, HIF-1, NF- and kappa B protein in 50 cases of cervical cancer, 40 cases of CIN and 26 cases of normal cervical tissue, and the relationship between the three groups was analyzed.
4. SPSS16.0 was used for statistical analysis, x 2 test and Spearman grade correlation analysis, the level of alpha =0.05 was tested, and the difference was statistically significant with the difference of P < 0.05.
Result
1. the expression level of IEX-1mRNA was significantly increased after different doses of UVB irradiation on human cervical cancer HeLa cells respectively (P0.05). The highest expression of IEX-1mRNA was observed after irradiation with UVB dose of 40mJ/cm2, which was the best ultraviolet radiation dose.
2., we irradiated human cervical cancer HeLa cells with the best ultraviolet dose of 40mJ/cm2, and detected the expression level of IEX-1mRNA after different time. After 4 hours of culture, the expression level of IEX-1mRNA was the highest (P0.05).
The positive expression of 3.IEX-1 in cervical cancer tissues was significantly lower than that in normal cervical tissue and cervical CIN, the difference was statistically significant (P0.05); the positive expression of HIF-1 kappa B alpha and NF- in cervical cancer tissues was significantly higher in cervical CIN and normal cervical tissue, the difference has statistical significance (P0.05).
4. the expression of IEX-1, HIF-1 and NF- kappa B in cervical cancer tissues was correlated with histological grade and lymph node metastasis (P0.05), which was not related to the age of patients, histological type and FIGO stage of tumor (P0.05).
5. the expression of HIF-1 alpha and NF- kappa B in cervical cancer tissues was positively correlated (r=0.398, P0.05), and they were negatively correlated with the expression of IEX-1 (r1=-0.322, P0.05; r2=-0.517, P0.05).
conclusion
In 1. human cervical cancer HeLa cells, IEX-1 is radiation-induced, which can be induced by UVB and has time and dose dependence.
The low expression of 2.IEX-1 in cervical cancer tissues is negatively correlated with the expression of NF- kappa B and HIF-1 alpha, and the three together regulate the growth and apoptosis of tumor cells.

【學(xué)位授予單位】：鄭州大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2014
【分類號(hào)】：R737.33

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相關(guān)期刊論文 前1條

1 肖濤;朱武;姚茂金;方建珍;羅遠(yuǎn)明;曾偉;胡金璽;;三氧化二砷下調(diào)MG-63骨肉瘤細(xì)胞IEX-1基因表達(dá)的轉(zhuǎn)錄調(diào)控機(jī)制研究[J];生物化學(xué)與生物物理進(jìn)展;2008年07期

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