蛋白激酶D1沉默可降低人涎腺腺样囊性癌细胞ACC-2对紫杉醇的敏感度
发布时间：2018-11-07 10:58
【摘要】：目的研究蛋白激酶D(PKD)沉默对人涎腺腺样囊性癌细胞ACC-2细胞增殖、迁移、化疗药物敏感度及凋亡的影响。方法转染Control-sh RNA和PKD1-sh RNA质粒后,药物筛选出稳定转染的ACC-2细胞系,并用Western blot验证细胞中PKD1敲除效率;划痕实验检测PKD1敲除后细胞迁移能力改变;CCK-8法检测PKD1敲除后细胞增殖能力以及紫杉醇对细胞的半致死浓度变化;PI染色并用流式细胞仪检测紫杉醇处理并PKD1敲除后细胞凋亡情况变化。结果建立了PKD1基因沉默的稳定细胞株;相较于对照组,实验组PKD1-sh细胞的增殖能力和迁移能力没有显著变化,但紫杉醇半致死浓度增高,紫杉醇处理后的细胞凋亡率降低。结论 PKD1沉默降低了ACC-2细胞对紫杉醇的药物敏感度,抑制了紫杉醇引起的细胞凋亡。
[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.
【作者单位】： 四川大学华西口腔医院口腔疾病研究国家重点实验室;
【基金】：国家自然科学基金(81372892)
【分类号】：R739.87
本文编号：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.
【作者单位】： 四川大学华西口腔医院口腔疾病研究国家重点实验室;
【基金】：国家自然科学基金(81372892)
【分类号】：R739.87
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