發(fā)布時(shí)間：2018-08-09 20:24

【摘要】：目的：建立穩(wěn)定表達(dá)CYB5R2基因的鼻咽癌細(xì)胞株,觀察CYB5R2基因?qū)Ρ茄拾┘?xì)胞惡性腫瘤生物行為的影響,驗(yàn)證其在鼻咽癌中的抑癌特性,為尋找更多的更有效的鼻咽癌分子生物學(xué)標(biāo)記物及治療新靶點(diǎn)提供理論依據(jù)和奠定實(shí)驗(yàn)基礎(chǔ)。方法：我們利用從CYB5R2高表達(dá)的正常睪丸組織cDNA中擴(kuò)增的CDS序列,構(gòu)建CYB5R2基因真核表達(dá)載體；將其轉(zhuǎn)染入CYB5R2表達(dá)沉默的鼻咽癌細(xì)胞株,篩選鑒定出穩(wěn)定表達(dá)CYN5R2的鼻咽癌細(xì)胞株；通過生長抑制實(shí)驗(yàn),克隆形成實(shí)驗(yàn)、遷移運(yùn)動(dòng)試驗(yàn)、裸鼠體內(nèi)成瘤試驗(yàn),觀察CYB5R2對(duì)鼻咽癌細(xì)胞的增殖、侵襲以及成瘤能力的影響。結(jié)果：成功構(gòu)建CYB5R2真核表達(dá)載體并獲得穩(wěn)定表達(dá)CYB5R2的鼻咽癌細(xì)胞株HONE1和CNE2。相對(duì)于轉(zhuǎn)染的空載體的鼻咽癌細(xì)胞株,轉(zhuǎn)染CYB5R2的鼻咽癌細(xì)胞的增殖能力、侵襲能力及裸鼠體內(nèi)成瘤能力明顯降低,均具有統(tǒng)計(jì)學(xué)意義。結(jié)論：我們的研究首次證明CYB5R2在體內(nèi)外均可抑制鼻咽癌細(xì)胞的惡性生物學(xué)行為,是一個(gè)鼻咽癌相關(guān)的潛在腫瘤抑制基因。
[Abstract]:Objective: to establish a nasopharyngeal carcinoma cell line stably expressing CYB5R2 gene and observe the effect of CYB5R2 gene on the biological behavior of nasopharyngeal carcinoma (NPC) cells. In order to find more effective molecular biological markers of nasopharyngeal carcinoma and new therapeutic targets to provide theoretical basis and lay the experimental foundation. Methods: we constructed the eukaryotic expression vector of CYB5R2 gene by using the CDS sequence amplified from cDNA of normal testicular tissue with high expression of CYB5R2, transfected it into NPC cell line with CYB5R2 expression silencing, and screened and identified the NPC cell line expressing CYN5R2 stably. The effects of CYB5R2 on the proliferation, invasion and tumorigenesis of nasopharyngeal carcinoma cells were observed by growth inhibition test, clone formation test, migration exercise test and tumorigenesis test in nude mice. Results: CYB5R2 eukaryotic expression vector was successfully constructed and NPC cell lines HONE1 and CNE2 expressing CYB5R2 stably were obtained. Compared with nasopharyngeal carcinoma cell line transfected with empty vector, the proliferation, invasion and tumorigenic ability of CYB5R2 transfected nasopharyngeal carcinoma cell line were significantly lower than that of empty vector transfected nasopharyngeal carcinoma cell line. Conclusion: our results suggest that CYB5R2 can inhibit the malignant biological behavior of nasopharyngeal carcinoma cells in vivo and in vitro, and is a potential tumor suppressor gene associated with nasopharyngeal carcinoma.
【學(xué)位授予單位】：廣西醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2012
【分類號(hào)】：R739.63

【參考文獻(xiàn)】

中國期刊全文數(shù)據(jù)庫 前8條

1 賀修勝,肖志強(qiáng),陳主初,趙素萍,朱建華,賀智敏,李友軍,田芳,余艷輝;STGC3新基因的克隆及功能初步分析[J];癌癥;2004年10期

2 張三泉,彭宏,宋蘭英,李先茂,蔣會(huì)勇,姚開泰,趙彤;組織芯片技術(shù)檢測人鼻咽癌組織及細(xì)胞株KIAA1173基因的表達(dá)[J];癌癥;2005年11期

3 張三泉,彭宏,將會(huì)勇,胡海,張進(jìn)華,姚開泰,趙彤;人KIAA1173基因的克隆、轉(zhuǎn)染及生物學(xué)特性的實(shí)驗(yàn)研究[J];第一軍醫(yī)大學(xué)學(xué)報(bào);2005年10期

4 趙曉榮,鄧琳,翁新憲,顧煥華,卓纓,鄧錫云,曹亞;鼻咽癌細(xì)胞中外源性p16表達(dá)對(duì)CDK4,CyclinD1及pRb的影響[J];湖南醫(yī)科大學(xué)學(xué)報(bào);2000年05期

5 李杰恩,黃光武,農(nóng)輝圖,徐志文,唐安洲,鄺國乾,韋敏怡;微血管密度與鼻咽癌生物學(xué)行為的相關(guān)性[J];臨床耳鼻咽喉科雜志;1999年08期

6 黃國良,張更,朱進(jìn)偉,高妍;NADH-細(xì)胞色素b_5還原酶在甲狀腺過氧化氫生物合成中的作用[J];生理學(xué)報(bào);2002年04期

7 項(xiàng)一寧,張維元;鼻咽癌組織中p16基因的缺失、高甲基化和蛋白的表達(dá)及其臨床意義[J];中華病理學(xué)雜志;2005年06期

8 李智,任藝,林素暇,梁英杰,梁惠珍;Association of E-cadherin and β-catenin with metastasis in nasopharyngeal carcinoma[J];Chinese Medical Journal;2004年08期

，

本文編號(hào)：2175215