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

【摘要】：目的:初步研究BCL9基因在三種乳腺癌細胞株中的表達差異,分析三種不同乳腺癌細胞株細胞生物學特性:增殖、侵襲及遷移能力的差異及其意義,并進一步探討B(tài)CL9基因的沉默對乳腺癌細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胞株,采用實時熒光定量PCR法(Real-time PCR)和蛋白免疫印跡法(Western blot)檢測人乳腺癌細胞株MDA-MB-231、MCF-7和MDA-MB-453中BCL9的mRNA及蛋白表達水平,篩選出高表達BCL9的細胞株。2通過流式細胞術(shù)檢測細胞周期,計算增殖指數(shù),比較三種細胞株之間增殖能力。采用細胞劃痕實驗和Transwell細胞侵襲實驗比較三者細胞遷移和侵襲能力。3構(gòu)建BCL9慢病毒質(zhì)粒shRNA,針對高表達BCL9的細胞株做BCL9基因沉默實驗。4采用Transwell細胞小室以及細胞劃痕實驗進一步研究BCL9基因的敲除對乳腺癌細胞株侵襲及遷移能力的影響。結(jié)果:1 RT-PCR檢測細胞株BCL9的mRNA表達: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)計學意義。BCL9蛋白水平在細胞間表達量為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)計學意義;2流式細胞術(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)計學意義;Transwell細胞侵襲實驗:穿膜細胞數(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)計學意義;細胞劃痕實驗: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)計學意義。3針對人乳腺癌細胞株MDA-MB-231運用慢病毒細胞轉(zhuǎn)染技術(shù)進行BCL9基因沉默;4 Transwell細胞小室實驗細胞數(shù):空白組(420.50±3.704)/陰性對照組(417.00±5.310)實驗組(151.50±7.580)(P0.05),差異有統(tǒng)計學意義;細胞劃痕實驗劃痕愈合率:空白組(0.712±0.127)/陰性對照組(0.709±0.015)實驗組(0.382±0.012)(P0.05),差異有統(tǒng)計學意義。結(jié)論:1通過檢測三種人乳腺癌細胞株中BCL9基因及蛋白的表達水平,發(fā)現(xiàn)BCL9在三陰性乳腺癌細胞株MDA-MB-231細胞中表達水平較高。2通過分析三種不同表型乳腺癌細胞株,細胞的增殖、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胞侵襲及轉(zhuǎn)移新的分子靶點。
[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：1953870