西南大學生命科學學院教育部創(chuàng)新重點實驗室

使用zeta life，Advanced DNA RNA轉染試劑

高效率轉染HCT-116, SW480, HT-29人結直腸癌細胞

2020-4-28發(fā)表文章已見刊

發(fā)表文章轉染條件

A、HCT-116, SW480, HT-29人結直腸癌細胞

B、AURKA質粒

C、轉染細胞融合度50%

D、96孔板每孔使用0.5μg質粒DNA

6孔每孔使用10 ug質粒DNA

(注意：本實驗中用到的細胞密度、轉染質粒DNA用量不適用于LIPO3000/2000)

發(fā)表文章部分內容

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.