Preliminary Study On The Effect Of Cisplatin On Nephroblastoma Cell Line In Vitro

Objective To investigate the inhibitory effects of different concentrations of Cisplatin(DDP)on nephroblastoma strains,and provide preliminary work and theoretical basis for related animal experiments.Methods 1.CCK-8 method was used to detect cell proliferation activity: two cancer cells(SK-NEP-1/G401)were cultured in 96-well plates,and the cancer cells were divided into 6 groups according to the amount of cisplatin added,at 24,48 After 72 h,CCK-8 was added,and its optical density(OD value)at 450 nm was measured and statistical analysis was performed.2.Scratch test to detect cell migration ability: two cancer cells were cultured on a 24-well plate,and the cancer cells were divided into 6 groups according to the amount of cisplatin added,and the gun head was crossed perpendicularly to the ruler at 0,24,48 h observed cell migration,photographed cell migration,calculated scratch healing rate% =(0 h scratch area pixel-24,48 h scratch area pixel)/0 h scratch area pixel x100%,and statistics analysis3.Western blot(WB)measured BAX protein content: According to the amount of cisplatin added,the cancer cells were divided into 6 groups,total protein was extracted,and the content of Bax protein and β-Actin protein in each group was detected by Western blot.Calculate the relative amount of Bax protein and perform statistical analysisResults 1.At 24,48,72 h after inoculation of cells,SK-NEP-1/G401 cells without cisplatin group(0% group)had significantly higher OD value at A450 than cisplatin group(0.1%).,0.2%,0.4%,0.8%,and 1.6% groups)(p < 0.001),the maximum inhibitory concentration of cisplatin on SK-NEP-1/G401 proliferation ability was 0.2% to 0.4%.2.At 0,24,48 h after inoculation,cisplatin had significant differences in the scratch healing rate of SK-NEP-1/G401 cells at 24 h and 48h(p<0.001),and cisplatin to SK-NEP-1 The maximum inhibitory concentration of migration ability is 0.2% to 0.4%.3.Bax expression increased after cisplatin treatment,0.2% group and 0,4% group were significantly different from the control group(p<0.001),and cisplatin caused a significant increase of Bax protein in SK-NEP-1 at 0%.~0.2%;0.4% group was significantly different from the control group(p<0.001),0.2% group and 0.4% group were significantly different(p<0.001),and cisplatin caused a significant increase in Bax protein concentration in G401.%~0.4%.Conclusion 1.Cisplatin can inhibit the proliferation of SK-NEP-1/G401 tumor cells,and the maximum inhibitory concentration of cisplatin on SK-NEP-1/G401 is 0.2%-0.4%.2.Cisplatin can inhibit the migration of SK-NEP-1/G401 tumor cells,and the maximum inhibitory concentration of cisplatin on SK-NEP-1 migration is 0.2%-0.4%.3.Cisplatin can promote the expression of Bax protein,which may be the reason for the induction of apoptosis and the ability of cancer to decrease.The concentration of Bax protein in SK-NEP-1 caused by cisplatin is 0%-0.2%,which is caused by cisplatin.The concentration of Bax protein in G401 was significantly increased from 0.2% to 0.4%.4.The inhibition of cisplatin on the nephroblastoma SK-NEP-1/G401 tumor strain exists in the plateau...