蛋白激酶D1沉默可降低人涎腺腺樣囊性癌細胞ACC-2對紫杉醇的敏感度
發(fā)布時間：2018-11-07 10:58
【摘要】：目的研究蛋白激酶D(PKD)沉默對人涎腺腺樣囊性癌細胞ACC-2細胞增殖、遷移、化療藥物敏感度及凋亡的影響。方法轉(zhuǎn)染Control-sh RNA和PKD1-sh RNA質(zhì)粒后,藥物篩選出穩(wěn)定轉(zhuǎn)染的ACC-2細胞系,并用Western blot驗證細胞中PKD1敲除效率;劃痕實驗檢測PKD1敲除后細胞遷移能力改變;CCK-8法檢測PKD1敲除后細胞增殖能力以及紫杉醇對細胞的半致死濃度變化;PI染色并用流式細胞儀檢測紫杉醇處理并PKD1敲除后細胞凋亡情況變化。結(jié)果建立了PKD1基因沉默的穩(wěn)定細胞株;相較于對照組,實驗組PKD1-sh細胞的增殖能力和遷移能力沒有顯著變化,但紫杉醇半致死濃度增高,紫杉醇處理后的細胞凋亡率降低。結(jié)論 PKD1沉默降低了ACC-2細胞對紫杉醇的藥物敏感度,抑制了紫杉醇引起的細胞凋亡。
[Abstract]:Objective to study the effects of protein kinase D (PKD) silencing on the proliferation, migration, chemotherapeutic sensitivity and apoptosis of human salivary adenoid cystic carcinoma ACC-2 cells. Methods after transfection of Control-sh RNA and PKD1-sh RNA plasmids, stably transfected ACC-2 cell lines were screened, and the PKD1 knockout efficiency was verified by Western blot, and the migration ability of PKD1 knockout cells was detected by scratch test. CCK-8 assay was used to detect the cell proliferation ability after PKD1 knockout and the change of half-lethal concentration of paclitaxel to cells, and PI staining and flow cytometry were used to detect apoptosis after paclitaxel treatment and PKD1 knockout. Results A stable cell line with PKD1 gene silencing was established, and the proliferation and migration ability of PKD1-sh cells in the experimental group was not significantly changed compared with the control group, but the half-lethal concentration of paclitaxel increased and the apoptosis rate decreased after paclitaxel treatment. Conclusion PKD1 silencing reduces the drug sensitivity of ACC-2 cells to paclitaxel and inhibits the apoptosis induced by paclitaxel.
【作者單位】： 四川大學華西口腔醫(yī)院口腔疾病研究國家重點實驗室;
【基金】：國家自然科學基金(81372892)
【分類號】：R739.87
本文編號：2316141
[Abstract]:Objective to study the effects of protein kinase D (PKD) silencing on the proliferation, migration, chemotherapeutic sensitivity and apoptosis of human salivary adenoid cystic carcinoma ACC-2 cells. Methods after transfection of Control-sh RNA and PKD1-sh RNA plasmids, stably transfected ACC-2 cell lines were screened, and the PKD1 knockout efficiency was verified by Western blot, and the migration ability of PKD1 knockout cells was detected by scratch test. CCK-8 assay was used to detect the cell proliferation ability after PKD1 knockout and the change of half-lethal concentration of paclitaxel to cells, and PI staining and flow cytometry were used to detect apoptosis after paclitaxel treatment and PKD1 knockout. Results A stable cell line with PKD1 gene silencing was established, and the proliferation and migration ability of PKD1-sh cells in the experimental group was not significantly changed compared with the control group, but the half-lethal concentration of paclitaxel increased and the apoptosis rate decreased after paclitaxel treatment. Conclusion PKD1 silencing reduces the drug sensitivity of ACC-2 cells to paclitaxel and inhibits the apoptosis induced by paclitaxel.
【作者單位】： 四川大學華西口腔醫(yī)院口腔疾病研究國家重點實驗室;
【基金】：國家自然科學基金(81372892)
【分類號】：R739.87
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