本文選題：BCAS2　切入點(diǎn)：顆粒細(xì)胞　出處：《華中農(nóng)業(yè)大學(xué)》2015年碩士論文

【摘要】：BCAS2是雌激素受體的一種輔助轉(zhuǎn)錄激活因子,其表達(dá)量的變化會(huì)影響雌激素受體相關(guān)基因的轉(zhuǎn)錄活性。雌激素在哺乳動(dòng)物卵泡發(fā)育中有重要的調(diào)控作用。目前關(guān)于BCAS2的研究主要集中在癌細(xì)胞系中,如乳腺癌細(xì)胞、Hela細(xì)胞等,而關(guān)于其在正常體細(xì)胞和生殖細(xì)胞內(nèi)的功能研究報(bào)道還較少。因此,本研究以小鼠為實(shí)驗(yàn)動(dòng)物模型,利用細(xì)胞免疫熒光、Western blot、Real-time PCR等實(shí)驗(yàn)技術(shù),檢測(cè)了BCAS2在小鼠卵巢組織中的定位和表達(dá);利用RNA干擾技術(shù)對(duì)BCAS2在小鼠卵巢顆粒細(xì)胞內(nèi)的功能做了初步的探索,結(jié)果如下:(1)BCAS2在小鼠顆粒細(xì)胞和卵母細(xì)胞中的表達(dá)和定位。細(xì)胞免疫熒光結(jié)果顯示,BCAS2主要定位在顆粒細(xì)胞和卵母細(xì)胞的細(xì)胞核中。Western blot結(jié)果顯示,BCAS2在小鼠GV-MII期卵母細(xì)胞中的表達(dá)量逐步升高。在超數(shù)排卵處理后,PMSG注射后6 h和12 h卵巢中,BCAS2的表達(dá)水平顯著高于其它時(shí)期(P0.05)。Real-time PCR結(jié)果顯示,BCAS2 m RNA在腔前卵泡中的表達(dá)量顯著高于有腔卵泡(P0.05)。FSH可以促進(jìn)體外培養(yǎng)顆粒細(xì)胞中BCAS2 m RNA的表達(dá)。(2)RNAi干擾敲低BCAS2的表達(dá)對(duì)顆粒細(xì)胞增殖和凋亡的影響。在顆粒細(xì)胞中干擾BCAS2后,能顯著性地降低了顆粒細(xì)胞的增殖水平(P0.05),同時(shí)顯著地下調(diào)增殖相關(guān)基因PCNA的表達(dá)(P0.05),干擾BCAS2之后,促進(jìn)顆粒細(xì)胞凋亡,并且抗凋亡基因BCL-2表達(dá)量顯著下調(diào),促凋亡基因BAX的表達(dá)量顯著上調(diào)(P0.05)。綜上,本研究發(fā)現(xiàn)BCAS2定位于小鼠顆粒細(xì)胞和卵母細(xì)胞的細(xì)胞核內(nèi),同時(shí),BCAS2在不同發(fā)育時(shí)期的卵泡顆粒細(xì)胞中具有特殊的表達(dá)規(guī)律,顆粒細(xì)胞中干擾BCAS2之后,會(huì)顯著降低細(xì)胞的增殖水平,并顯著促進(jìn)細(xì)胞凋亡,同時(shí),顯著性影響增殖和凋亡相關(guān)基因的表達(dá),該結(jié)果說(shuō)明BCAS2可能參與卵泡顆粒細(xì)胞增殖和凋亡的生理過程。
[Abstract]:BCAS2 is an assistant transcriptional activator of estrogen receptor, and its expression will affect the transcriptional activity of estrogen receptor related genes.Estrogen plays an important role in the regulation of follicle development in mammals.At present, the study of BCAS2 is mainly focused on cancer cell lines, such as breast cancer cells, such as Hela cells, but its function in normal somatic cells and germ cells is less reported.Therefore, in this study, the localization and expression of BCAS2 in mouse ovarian tissues were detected by using CIFA Western blotreal-time PCR and other experimental techniques.The function of BCAS2 in mouse ovarian granulosa cells was studied by RNA interference technique. The results were as follows: the expression and localization of BCAS2 in mouse granulosa cells and oocytes.The results of cellular immunofluorescence showed that BCAS2 was mainly located in the nucleus of granulosa cells and oocytes. Western blot showed that the expression of BCAS2 in mouse GV-MII oocytes increased gradually.Expression of BCAS2 m RNA in granulosa cells. Effect of RNAi interference knockout on proliferation and apoptosis of granulosa cells.After interfering with BCAS2 in granulosa cells, the proliferation level of granulosa cells was significantly decreased, and the expression of proliferation-related gene PCNA was significantly down-regulated. After interfering with BCAS2, granulosa cells apoptosis was promoted.The expression of anti-apoptotic gene BCL-2 was down-regulated, and the expression of pro-apoptotic gene BAX was significantly up-regulated (P 0.05).In conclusion, we found that BCAS2 was located in the nucleus of mouse granulosa cells and oocytes, and that BCAS2 had a special expression pattern in follicular granulosa cells at different developmental stages, after interfering with BCAS2 in granulosa cells.The results indicated that BCAS2 may be involved in the physiological process of proliferation and apoptosis of follicular granulosa cells.
【學(xué)位授予單位】：華中農(nóng)業(yè)大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2015
【分類號(hào)】：S814

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