本文選題：乳腺癌 + B細(xì)胞淋巴因子9　； 參考：《河北醫(yī)科大學(xué)》2017年碩士論文

【摘要】：目的:初步研究BCL9基因在三種乳腺癌細(xì)胞株中的表達(dá)差異,分析三種不同乳腺癌細(xì)胞株細(xì)胞生物學(xué)特性:增殖、侵襲及遷移能力的差異及其意義,并進(jìn)一步探討B(tài)CL9基因的沉默對(duì)乳腺癌細(xì)胞株侵襲及遷移能力的影響及意義。方法:1分析三種不同表型的人乳腺癌細(xì)胞株,采用實(shí)時(shí)熒光定量PCR法(Real-time PCR)和蛋白免疫印跡法(Western blot)檢測(cè)人乳腺癌細(xì)胞株MDA-MB-231、MCF-7和MDA-MB-453中BCL9的mRNA及蛋白表達(dá)水平,篩選出高表達(dá)BCL9的細(xì)胞株。2通過(guò)流式細(xì)胞術(shù)檢測(cè)細(xì)胞周期,計(jì)算增殖指數(shù),比較三種細(xì)胞株之間增殖能力。采用細(xì)胞劃痕實(shí)驗(yàn)和Transwell細(xì)胞侵襲實(shí)驗(yàn)比較三者細(xì)胞遷移和侵襲能力。3構(gòu)建BCL9慢病毒質(zhì)粒shRNA,針對(duì)高表達(dá)BCL9的細(xì)胞株做BCL9基因沉默實(shí)驗(yàn)。4采用Transwell細(xì)胞小室以及細(xì)胞劃痕實(shí)驗(yàn)進(jìn)一步研究BCL9基因的敲除對(duì)乳腺癌細(xì)胞株侵襲及遷移能力的影響。結(jié)果:1 RT-PCR檢測(cè)細(xì)胞株BCL9的mRNA表達(dá):MDA-MB-231(0.016±0.004)MCF-7(0.008±0.002)MDA-MB-453(0.004±0.002),(各組間比較,P0.05)差異有統(tǒng)計(jì)學(xué)意義。BCL9蛋白水平在細(xì)胞間表達(dá)量為MDA-MB-231(0.629±0.101)MCF-7(0.397±0.196)MDA-MB-453(0.204±0.990)(各組間比較,P0.05),差異有統(tǒng)計(jì)學(xué)意義;2流式細(xì)胞術(shù)增殖指數(shù)顯示MDA-MB-231(0.483±0.010)MDA-MB-453(0.358±0.032),MCF-7(0.397±0.061)(各組間比較,P0.05),差別有統(tǒng)計(jì)學(xué)意義;Transwell細(xì)胞侵襲實(shí)驗(yàn):穿膜細(xì)胞數(shù):MDA-MB-231(417.80±6.94)MDA-MB-453(379.60±11.99)MCF-7(9.40±1.14)(各組間比較,P0.05),差異有統(tǒng)計(jì)學(xué)意義;細(xì)胞劃痕實(shí)驗(yàn):MDA-MB-231(0.716±0.021)MDA-MB-453(0.331±0.021)MCF-7(0.272±0.017)(各組間比較,P0.05),差異有統(tǒng)計(jì)學(xué)意義。3針對(duì)人乳腺癌細(xì)胞株MDA-MB-231運(yùn)用慢病毒細(xì)胞轉(zhuǎn)染技術(shù)進(jìn)行BCL9基因沉默;4 Transwell細(xì)胞小室實(shí)驗(yàn)細(xì)胞數(shù):空白組(420.50±3.704)/陰性對(duì)照組(417.00±5.310)實(shí)驗(yàn)組(151.50±7.580)(P0.05),差異有統(tǒng)計(jì)學(xué)意義;細(xì)胞劃痕實(shí)驗(yàn)劃痕愈合率:空白組(0.712±0.127)/陰性對(duì)照組(0.709±0.015)實(shí)驗(yàn)組(0.382±0.012)(P0.05),差異有統(tǒng)計(jì)學(xué)意義。結(jié)論:1通過(guò)檢測(cè)三種人乳腺癌細(xì)胞株中BCL9基因及蛋白的表達(dá)水平,發(fā)現(xiàn)BCL9在三陰性乳腺癌細(xì)胞株MDA-MB-231細(xì)胞中表達(dá)水平較高。2通過(guò)分析三種不同表型乳腺癌細(xì)胞株,細(xì)胞的增殖、侵襲和遷移能力均有差別,三陰性乳腺癌的MDA-MB-231細(xì)胞株的增殖、侵襲和遷移能力均較高。3 BCL9基因的沉默可有效抑制人乳腺癌細(xì)胞株MDA-MB-231侵襲及遷移能力,提示BCL9可能成為抑制乳腺癌細(xì)胞侵襲及轉(zhuǎn)移新的分子靶點(diǎn)。
[Abstract]:Objective: to study the difference of BCL9 gene expression in three breast cancer cell lines, and analyze the biological characteristics of three different breast cancer cell lines: proliferation, invasion and migration. The effect and significance of BCL9 gene silencing on the invasion and migration of breast cancer cell lines were investigated. Methods three human breast cancer cell lines with different phenotypes were analyzed. The mRNA and protein expression of BCL9 in MDA-MB-231 MCF-7 and MDA-MB-453 were detected by real-time fluorescence quantitative PCR and Western blot. The cell lines with high expression of BCL9 were selected. The cell cycle was detected by flow cytometry, the proliferation index was calculated, and the proliferative ability of the three cell lines was compared. Cell scrape assay and Transwell cell invasion assay were used to compare the cell migration and invasion ability of three cells to construct BCL9 lentivirus plasmid shRNAs. The BCL9 gene silencing assay was performed on the cell lines with high expression of BCL9. 4. Transwell cell compartment and cell division were used. The effect of BCL9 knockout on the invasion and migration of breast cancer cell lines was further studied. 緇撴灉:1 RT-PCR媯，

本文編號(hào)：1953870