發(fā)布時(shí)間：2018-03-12 23:12

  本文選題：宮頸癌　切入點(diǎn)：放療敏感性　出處：《南方醫(yī)科大學(xué)》2017年碩士論文　論文類型：學(xué)位論文

【摘要】：研究背景和目的宮頸癌是女性生殖系統(tǒng)中最常見(jiàn)的惡性腫瘤。而放療是宮頸癌的治療手段之一,80%患者需接受放療,而30-50%的晚期患者出現(xiàn)了放療抵抗。因此,提高放療敏感性對(duì)晚期宮頸癌患者而言十分重要。如今對(duì)宮頸癌的研究發(fā)現(xiàn)一些癌基因、轉(zhuǎn)錄因子等與宮頸癌的放療敏感性密切相關(guān),但仍缺乏公認(rèn)的分子標(biāo)志物。因此,尋找相關(guān)分子并研究其調(diào)控機(jī)制,對(duì)改善宮頸癌放療敏感具有重要的意義。轉(zhuǎn)錄調(diào)控因子Sp1是一個(gè)具有序列特異性的DNA結(jié)合蛋白,參與調(diào)控了一些腫瘤的放療敏感性,且其與宮頸癌的進(jìn)展密切相關(guān)。因此,Sp1可能是影響宮頸癌放療敏感的一個(gè)重要因子。本研究發(fā)現(xiàn)放療可上調(diào)宮頸癌細(xì)胞中Sp1的表達(dá),通過(guò)沉默/過(guò)表達(dá)SiHa、HeLa中Sp1的表達(dá),揭示了 Sp1通過(guò)上調(diào)CDK1的表達(dá)減少放療相關(guān)的G2/M期阻滯從而下調(diào)宮頸癌的放療敏感性。本研究將分四部分進(jìn)行。第一章Sp1與宮頸癌放療的相關(guān)性研究目的研究Sp1與宮頸癌放療的相關(guān)性方法利用western blot和RT-PCR檢測(cè)放療后宮頸癌細(xì)胞中Sp1的變化。結(jié)果宮頸癌細(xì)胞中Sp1的表達(dá)水平隨照射劑量的增加而增加。結(jié)論Sp1與宮頸癌放療密切相關(guān)。第二章Sp1與宮頸癌細(xì)胞放療敏感性的關(guān)系目的探討Sp1對(duì)宮頸癌放療敏感性的影響方法構(gòu)建Sp1沉默/過(guò)表達(dá)的宮頸癌細(xì)胞株,利用克隆形成及CCK8實(shí)驗(yàn)檢測(cè)Sp1對(duì)宮頸癌放療敏感性的影響。結(jié)果沉默Sp1可顯著上調(diào)宮頸癌的放療敏感性,過(guò)表達(dá)Sp1則相反。結(jié)論Sp1可下調(diào)宮頸癌的放療敏感性。第三章Sp1通過(guò)調(diào)控放療相關(guān)的細(xì)胞周期阻滯影響宮頸癌細(xì)胞放療敏感性目的研究Sp1對(duì)宮頸癌細(xì)胞周期的影響方法利用流式細(xì)胞術(shù)檢測(cè)Sp1對(duì)宮頸癌放療相關(guān)細(xì)胞周期阻滯的影響結(jié)果抑制Sp1的表達(dá)可顯著上調(diào)宮頸癌的G2/M期阻滯(p0.05),而過(guò)表達(dá)Sp1則顯著減少宮頸癌的G2/M期阻滯(p0.05)結(jié)論Sp1可通過(guò)下調(diào)放療相關(guān)的G2/M期阻滯從而下調(diào)宮頸癌的放療敏感性。第四章Sp1通過(guò)調(diào)控CDK1影響宮頸癌細(xì)胞的放療敏感性目的研究Sp1影響宮頸癌放療敏感性的分子機(jī)制。方法1.利用實(shí)時(shí)熒光定量PCR及蛋白免疫印跡檢測(cè)沉默/過(guò)表達(dá)Sp1后G2/M周期相關(guān)的基因變化2.利用恢復(fù)試驗(yàn)驗(yàn)證Sp1通過(guò)調(diào)控CDK1影響宮頸癌的放療敏感性3.利用雙熒光素酶實(shí)驗(yàn)驗(yàn)證Sp1是否作用于CDK1的啟動(dòng)子結(jié)果1.Sp1可上調(diào)CDK1的表達(dá)(P0.05)2.下調(diào)CDK1的表達(dá)后,Sp1對(duì)宮頸癌放療敏感性的下調(diào)作用減小(P0.05)3.Sp1顯著上調(diào)CDK1啟動(dòng)子的熒光活性(P0.01)結(jié)論Sp1可通過(guò)上調(diào)CDK1的表達(dá)下調(diào)宮頸癌的放療敏感性。全文總結(jié)1.Sp1與宮頸癌放療敏感性密切相關(guān),沉默/過(guò)表達(dá)Sp1可顯著上調(diào)/下調(diào)宮頸癌的放療敏感性2.Sp1通過(guò)減少輻射相關(guān)的G2/M期阻滯下調(diào)宮頸癌的放療敏感性3.Sp1通過(guò)上調(diào)CDK1的表達(dá)下調(diào)宮頸癌的放療敏感性本項(xiàng)目的創(chuàng)新之處1.驗(yàn)證了 Sp1對(duì)宮頸癌放療敏感性的作用;2.揭示Sp1通過(guò)調(diào)控CDK1下調(diào)宮頸癌放療敏感性的分子機(jī)制。
[Abstract]:Background and objective Cervical cancer is the most common malignant tumor in the female reproductive system. Radiotherapy is one of the treatments for cervical cancer. 80% of the patients need radiotherapy, while 30-50% of the advanced patients have radiation resistance. Increasing radiosensitivity is important for patients with advanced cervical cancer. Research on cervical cancer has found that some oncogenes, transcription factors, and so on are closely related to the radiosensitivity of cervical cancer, but still lack of recognized molecular markers. It is important to search for the related molecules and study its regulatory mechanism to improve the radiosensitivity of cervical cancer. The transcription regulator Sp1 is a sequence-specific DNA binding protein involved in regulating the radiosensitivity of some tumors. Sp1 may be an important factor affecting the sensitivity of cervical cancer to radiotherapy. In this study, we found that radiotherapy can up-regulate the expression of Sp1 in cervical cancer cells, and by silencing / overexpressing the expression of Sp1 in SiHa-HeLa, it may be an important factor affecting the radiosensitivity of cervical cancer. It is revealed that Sp1 down-regulates the radiosensitivity of cervical cancer by up-regulating the expression of CDK1 and reducing the radiotherapy-associated G _ 2 / M block. This study will be conducted in four parts. Chapter 1: study on the correlation between Sp1 and radiotherapy for cervical cancer objective to study Sp1. Methods western blot and RT-PCR were used to detect the changes of Sp1 in cervical cancer cells after radiotherapy. Results the expression of Sp1 in cervical cancer cells increased with the increase of irradiation dose. Conclusion Sp1 is closely associated with radiotherapy for cervical cancer. Chapter 2 relationship between Sp1 and radiosensitivity of cervical cancer cells objective to investigate the effects of Sp1 on the radiosensitivity of cervical cancer cells. The effects of Sp1 on the radiosensitivity of cervical cancer were detected by clone formation and CCK8 assay. Results silencing Sp1 could significantly up-regulate the radiosensitivity of cervical cancer. Conclusion Sp1 can down-regulate the radiosensitivity of cervical cancer cells by overexpression of Sp1. Chapter 3: to study the effect of Sp1 on the radiosensitivity of cervical cancer cells by regulating the cell cycle block associated with radiotherapy objective to study the effect of Sp1 on the radiosensitivity of cervical cancer cells. Methods flow cytometry was used to detect the effect of Sp1 on cell cycle arrest associated with radiotherapy for cervical cancer. Inhibiting the expression of Sp1 could significantly up-regulate the G _ 2 / M phase block of cervical cancer, while overexpression of Sp1 significantly decreased the G _ 2 / M phase block of cervical cancer. Conclusion Sp1 can down-regulate the radiosensitivity of cervical cancer by down-regulating the G _ 2 / M phase block associated with radiotherapy. Chapter 4th Sp1 studies the effects of Sp1 on the radiosensitivity of cervical cancer by regulating the radiosensitivity of cervical cancer cells by regulating CDK1. Methods 1. Real-time fluorescent quantitative PCR and Western blotting were used to detect the G 2 / M cycle related gene changes after silencing / overexpression of Sp1 2.Using recovery test to verify the effect of Sp1 on the radiosensitivity of cervical cancer by regulating CDK1. 2. 3. Double luciferase assay was used to verify whether Sp1 acted on the promoter of CDK1. 1. Sp1 could up-regulate the expression of CDK1 and P0.05 ~ 2. After down-regulating the expression of CDK1, the down-regulation of pSp1 on radiosensitivity of cervical cancer decreased P0.05 ~ (3) Sp1 significantly up-regulated the fluorescence of CDK1 promoter. Conclusion Sp1 can down-regulate the radiosensitivity of cervical cancer by up-regulating the expression of CDK1. 1. Sp1 is closely related to the radiosensitivity of cervical cancer. Silencing / overexpression of Sp1 could significantly up-regulate / down-regulate the radiosensitivity of cervical cancer 2.Sp1 down-regulates the radiosensitivity of cervical cancer by reducing radiation-related G _ 2 / M block 3.Sp1 down-regulates the radiosensitivity of cervical cancer by up-regulating the expression of CDK1. The role of Sp1 in the radiosensitivity of cervical cancer 2.The molecular mechanism of Sp1 down-regulating the radiosensitivity of cervical cancer by regulating CDK1. 2.
【學(xué)位授予單位】：南方醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R737.33

【參考文獻(xiàn)】

相關(guān)期刊論文 前1條

1 高燕,林莉萍,丁健;細(xì)胞周期調(diào)控的研究進(jìn)展[J];生命科學(xué);2005年04期

，

本文編號(hào)：1603734