西南大学高效率转染HCT116,SW480,HT29人结直肠癌细胞案例

西南大学生命科学学院教育部创新重点实验室

使用zeta life，Advanced DNA RNA转染试剂

高效率转染HCT-116, SW480, HT-29人结直肠癌细胞

2020-4-28发表文章已见刊

发表文章转染条件

A、HCT-116, SW480, HT-29人结直肠癌细胞

B、AURKA质粒

C、转染细胞融合度50%

D、96孔板每孔使用0.5μg质粒DNA

6孔每孔使用10 ug质粒DNA

(注意：本实验中用到的细胞密度、转染质粒DNA用量不适用于LIPO3000/2000)

发表文章部分内容

Overexpression of AURKA Plasmid for AURKA overexpression was obtained from VectorBuilder. Details about plasmid containing the AURKA gene can be found at vectorbuilder/vector/VB190802-1063ncc. html. colon cancer cells (HCT-116, SW480, HT-29) were seeded at 50% confluency for 24 h in six-well and 96-well plates. Plasmid (10 ug per-well for six-well and 0.5 μg per-well for 96 plates) and the Advanced DNA RNA transfection（zeta life,USA) reagent were mixed in equal proportionsand added to the cell culture medium for 6 h. DOX (1 μg/ml) was added to induce the expression of AURKA. After addition of DOX and PAL for 36 h, cell viability was measured by MTT and apoptosis was detected by。

PAL exerts anti-colon cancer effects by targeting AURKA. (A) PAL promotes apoptosis in colon cancer cells (HCT-116, SW480) in a dose-dependent manner. (B) PAL promotes G2/M phase arrest in colon cancer cells (HCT-116, SW480) in a dose-dependent manner. (C) The expression of AURKA after DOX treatment for 24 h. (D) Cell density after PAL treatment for 24 h in the cells (HCT-116, SW480) transfected with AURKA overexpression plasmids. DOX is an inducer of AURKA expression in plasmids. (E) Effect of PAL detected by MTT assay on the proliferation ability of colon cancer cells transfected with AURKA overexpression plasmids. (F) Effects of PAL detected by.