本文選題：Cyclin + D1�。� 參考：《吉林大學(xué)》2006年碩士論文

【摘要】：細(xì)胞周期蛋白D1(Cyclin D1)是一類重要的細(xì)胞周期正性調(diào)控因子,它通過(guò)激活CDK4/6來(lái)推進(jìn)細(xì)胞周期G1期到S期的演進(jìn)。Cyclin D1在很多惡性腫瘤中呈過(guò)度表達(dá),并與腫瘤的發(fā)生、惡性程度與患者的預(yù)后密切相關(guān)。Cyclin D1成為人們進(jìn)行腫瘤基因治療的靶點(diǎn)之一。 研究者們?cè)鴩L試向腫瘤細(xì)胞中注射抗Cyclin D1抗體、轉(zhuǎn)錄導(dǎo)入反義Cyclin D1,兩種方法均可改變轉(zhuǎn)化細(xì)胞形態(tài),抑制腫瘤細(xì)胞增殖,但是由于存在免疫原性較強(qiáng),后者又存在體內(nèi)半衰期短、不穩(wěn)定等缺點(diǎn),限制了該法在腫瘤治療中的應(yīng)用。近年來(lái)興起的以抗體作為效應(yīng)分子的能夠在細(xì)胞內(nèi)高效、特異滅活靶蛋白的胞內(nèi)抗體技術(shù)為腫瘤的基因治療開(kāi)辟了新的思路。 為此,本研究克隆了內(nèi)質(zhì)網(wǎng)滯留型抗人Cyclin D1胞內(nèi)scFv基因(ER-ADx),并構(gòu)建其重組真核表達(dá)載體pER-ADx,通過(guò)細(xì)胞轉(zhuǎn)染實(shí)驗(yàn)、RT-PCR、免疫組化、Dot-blot、MTT法生長(zhǎng)增殖特性分析、流式細(xì)胞術(shù)等方法和技術(shù)分別對(duì)ER-ADx的表達(dá)及胞內(nèi)定位情況、體外生物活性等進(jìn)行了研究。結(jié)果表明,ER-ADx基因能夠在腫瘤細(xì)胞中有效表達(dá)并實(shí)現(xiàn)胞內(nèi)抗體的目標(biāo)定位。ER-ADx基因的穩(wěn)定表達(dá)能夠顯著抑制細(xì)胞生長(zhǎng)和增殖,引起細(xì)胞周期阻滯,并明顯誘導(dǎo)細(xì)胞凋亡。綜上所述,以Cyclin D1為靶點(diǎn)的胞內(nèi)抗體治療作為一種新型腫瘤基因治療方法,有潛在的應(yīng)用價(jià)值。
[Abstract]:Cyclin D1 (cyclin D1) is an important positive regulator of cell cycle, which activates CDK4/6 to promote the progression from G1 to S phase of cell cycle. Cyclin D1 is overexpressed in many malignant tumors and is associated with tumorigenesis. The degree of malignancy is closely related to the prognosis of patients. Cyclin D1 has become one of the targets of gene therapy for cancer. The researchers have tried to inject anti-Cyclin D1 antibodies into tumor cells and transcribe them into antisense Cyclin D1. Both methods can change the morphology of transformed cells and inhibit the proliferation of tumor cells, but because of their strong immunogenicity, The latter has the shortcomings of short half-life and instability in vivo, which limits the application of this method in tumor therapy. In recent years, the technology of intracellular antibody with antibody as effector molecule can be highly efficient and specific to kill target protein in cells, which opens a new way for gene therapy of tumor. The recombinant eukaryotic expression vector pER-ADX was constructed by cloning endoplasmic reticulum (ER) retained anti-human Cyclin D1 scFv gene (ER-ADxX), and the proliferation characteristics of the recombinant plasmid pER-ADX were analyzed by cell transfection assay with RT-PCR and immunohistochemistry with Dot-blot MTT assay. The expression and intracellular localization of ER-ADx and its bioactivity in vitro were studied by flow cytometry. The results showed that ER-ADX gene could effectively express in tumor cells and realize the target localization of intracellular antibody. The stable expression of ER-ADX gene could significantly inhibit cell growth and proliferation, induce cell cycle arrest and induce apoptosis. In conclusion, intracellular antibody therapy targeting Cyclin D1 as a new tumor gene therapy has potential application value.
【學(xué)位授予單位】：吉林大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2006
【分類號(hào)】：R730.5;R392

【引證文獻(xiàn)】

相關(guān)碩士學(xué)位論文 前3條

1 劉志強(qiáng);抗Cyclin D1人源胞內(nèi)單鏈抗體的原核表達(dá)與鑒定[D];吉林大學(xué);2011年

2 李兵直;抗Cyclin D1胞內(nèi)抗體抑制腫瘤生長(zhǎng)和轉(zhuǎn)移[D];吉林大學(xué);2012年

3 王天時(shí);抗Cyclin D1胞內(nèi)抗體分子調(diào)控機(jī)制的蛋白質(zhì)組學(xué)研究[D];吉林大學(xué);2013年

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本文編號(hào)：1913856