西南大學(xué)高效率轉(zhuǎn)染HCT116,SW480,HT29人結(jié)直腸癌細(xì)胞案例

西南大學(xué)生命科學(xué)學(xué)院教育部創(chuàng)新重點實驗室

使用zeta life，Advanced DNA RNA轉(zhuǎn)染試劑

高效率轉(zhuǎn)染HCT-116, SW480, HT-29人結(jié)直腸癌細(xì)胞

2020-4-28發(fā)表文章已見刊

發(fā)表文章轉(zhuǎn)染條件

A、HCT-116, SW480, HT-29人結(jié)直腸癌細(xì)胞

B、AURKA質(zhì)粒

C、轉(zhuǎn)染細(xì)胞融合度50%

D、96孔板每孔使用0.5μg質(zhì)粒DNA

6孔每孔使用10 ug質(zhì)粒DNA

(注意：本實驗中用到的細(xì)胞密度、轉(zhuǎn)染質(zhì)粒DNA用量不適用于LIPO3000/2000)

發(fā)表文章部分內(nèi)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.