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Several anticancer drugs induced apoptosis of Jurkat cells
Chinese papers League finishing. Abstract The clinical evaluation of several commonly used anticancer drug cisplatin (CDDP), hydroxycamptothecin (HCPT), homoharringtonine base ( HHT) and mitoxantrone (MIT) in the treatment of leukemia and to analyze its role in inducing apoptosis of Jurkat cells for the clinical treatment of leukemia, a new theoretical basis and ideas. With Annexin V / PI dual parameter flow analysis to detect these types of Jurkat cell apoptosis induced by anticancer drugs and death effects. The results showed that: MIT and the HCPT-treatment in the early 4 hours (P Key words anti-cancer drugs; apoptosis; Jurkat cell line; Annexin V Effect of Several Anti-tumor Drugs on Apoptosis Induction in Jurkat Cell Line Abstract Cisplatin (CDDP), homoharringtonine (HHT), mitoxantrone (MIT) and hydroxycamptothecin (HCPT) are highly effective anti-tumor drugs. To evaluate thEir effects in the therapy of leukemia and establish a valuable method to estimate anti-tumor drugs, Annexin V / PI double parameter flow cytometry was used to detect the effects of these drug inducing apoptosis and death in Jurkat cell line. The results showed that MIT and HCTP-induced apoptosis effects on Jurkat cell line were obvious at 4 hours in early phase after adding drug (P <0.05) and at 8 hours in late phase after adding drug (P <0.05). HHT had obvious effect on inducing apoptosis of Jurkat cells, but no significant difference from low to high doses. The effect of CDDP on inducing apoptosis of Jurkat cell line was obviously weaker than that of HHT, MIT and HCPT, its weak effect on apoptosis of Jurkat cell line was found only at high concentration of drug for long time. Death effects on Jurkat cell line can not be observed in every experimental group. It is concluded that low dose of MIT can effectively induced apoptosis of Jurkat cell line. Annexin V / PI double parameter flow cytometry can be used as a reliable method for clinical screening anti-tumor drugs. Key words anti-tumor drug; apoptosis; Jurkat cell line; Annexin V chemotherapy is an important method of cancer therapy, new anticancer drugs are constantly emerging . China in the last century 90 listing the chemical compound classes of anticancer drugs mitoxantrone (MIT), hydroxyurea and cisplatin (CDDP) and a few of several anti-cancer drugs, through inhibition of nucleic acid molecules with the DNA synthesis induced cell death Such drugs called non-cell cycle specific drugs. In this study mitoxantrone (MIT), cisplatin (CDDP),[link widoczny dla zalogowanych], hydroxycamptothecin (HCPT), alkali homoharringtonine (HHT) induced apoptosis in Jurkat cells, and by Annexin V / PI apoptosis assay and the percentage of cell death, of different time and different concentration of Jurkat cell apoptosis. Materials and methods material Jurkat cells kept in our laboratory. Cisplatin (CDDP) for the Qilu Pharmaceutical Co., Ltd., hydroxycamptothecin (HCPT) as a yellow Dan Feiyun pharmaceutical products, base homoharringtonine (HHT) to Hangzhou Minsheng Pharmaceutical Products,[link widoczny dla zalogowanych], mitoxantrone ( MIT) for the new pharmaceutical products in Zhejiang Rui; RPMI 1640 as Invitrgen products; Annexin V FITC / PI kit was purchased from Jingmei company; Beike Man flow cytometry Elite EXP. induced apoptosis in Jurkat cells Jurkat cells in 5% CO2 incubator culture to logarithmic phase, microscopic count of 1 × 106/ml , adding 24-well plate, 1 ml cell suspension per well. Serum-free cell suspension to cisplatin,[link widoczny dla zalogowanych], HCPT, homoharringtonine base and preparation of drugs mitoxantrone 1 mg / ml solution; each concentration of drugs were located 4: 12.5,25,50,100 μg / ml; were added to 24-well culture plates for three duplicate wells. Placed in 5% CO2 incubator culture. Annexin V / PI dual parameter flow analysis [1-3] drugs were added 2,4 and 8 hours,[link widoczny dla zalogowanych], drawing cultured cells suspension, PBS precooled at 4 ℃ Wash cells 2 times with 250 μl binding buffer, resuspended cells to a concentration of 1 × 106/ml; take 100 μl cell suspension, add 5 μl Annexin V / FITC and 10 μl (20 μg / ml) of propidium iodide; mixing at room temperature away from light for 15 minutes; washed 2 times with PBS, for flow cytometry (FACS) analysis. Annexin V-FITC/PI double-parameter for debugging, this condition undergo apoptosis and necrosis rate detection. Figure 1-5 that the fluorescence intensity of the abscissa, vertical axis cells, two peaks in the figure that the negative peak of the left peak and right peak that apoptosis peak, the larger the area under the curve of the right peak the more apoptosis. According to the measured data based on cell apoptosis and secondary necrosis. statistical analysis apoptosis rate of cells in each group the mean of samples from 3 to X ± SD, said several of the two numbers used to compare the two, Results CDDP on induction of apoptosis in Jurkat cells CDDP dose group and the control of the group (untreated group) than in the dose of 12.5 μg / ml, the effect of time on the various Jurkat cell apoptosis rate was no significant change; dose of 25 μg / ml and 50 μg / ml,[link widoczny dla zalogowanych], in the two hours and 4 hours Jurkat cell apoptosis was also no significant change for 8 h in Jurkat cells have significantly higher rates were 13.6% and 23.9%, compared with the control group were significantly different (P <0.05) . When CDDP concentration increased to 100 μg / ml, the effect of 2 and 4 h, Jurkat cell apoptosis rate unchanged at 8 h was 19.9% in Jurkat cells with 50 μg / ml dose compared induced apoptosis has not significantly different, but decreased (Figure 1), suggesting that the role of CDDP with drug saturation effect. HHT induced apoptosis of Jurkat cells, the role of HHT dose group, the apoptosis rate with the control group (untreated group) were compared significantly higher (P <0.05). Jurkat cell apoptosis rate and no significant relationship between drug dose, but was proportional to the duration of action. 12.5 μg / ml HHT 2,4 and 8 hours when the role of Jurkat cell apoptosis rate was 4.9%, 20.4% and 35.4% (P <0.05); 25 μg / ml HHT 5.5%, respectively, 12.1% and 27.2% ; 50 μg / ml HHT 5.8%, respectively, 12.2% and 33.0%; 100 μg / ml HHT is 5.7%, respectively, 12.1% and 35.0% (P <0.05). Different drug concentrations at the same time point between HHT induced Jurkat cell apoptosis was no significant difference (Figure 2). MIT on Jurkat cell apoptosis induced by with the control group (untreated group) compared to, MIT in the effect of each dose group at 2 h Jurkat cells was not significantly different, but the action time with significantly higher (P <0.05). 12.5 μg / ml dose of 2,4 and 8 hours when the role of Jurkat cell apoptosis rate was 4.2%, 20.5% and 52.4% (P <0.05), 25 μg / ml group were 4.9%, 30.1% and 68.9% , 50 μg / ml group were 7.2%, 37.9% and 81.8% (P <0.05) (Figure 3), 100 μg / ml group were 6.6%, 41.5% and 84.1%, the effect of the differences between time points were There were significant differences (P <0.05) (Figure 4). Jurkat cell apoptosis in the drug dose increased with low doses significantly increased; 100 μg / ml group Effect and 50 μg / ml dose group, no significant difference, indicating a dose-saturation effect. HCPT induced apoptosis of Jurkat cells, the role of HCPT each dose group and control group (untreated group) compared to, Jurkat cell apoptosis significantly higher (P <0.05), Jurkat cell apoptosis increased with the dose and time extension was significantly higher, 12.5 μg / ml HCPT dose effect of 2,4 and 8 h Jurkat cell apoptosis rate was 3.4% , 20.9% and 30.3% (P <0.05); and 25 μg / ml group were 3.7%, 19.1% and 30.3%; 50 μg / ml group were 5.9%, 23.8% and 44.6% (P <0.05), 100 μg / ml group were 3.0%, 15.8% and 55.7% (P <0.05). HCPT different time points the same dose of Jurkat cell apoptosis rate was significantly different (P <0.05) (Figure 5); HCPT effect of different doses of the same time the induction of Jurkat cell apoptosis in 2 hours and 4 hours time was not significantly differences, both with the 8 hours were significantly different compared with the role (P <0.05). each drug group compared induced apoptosis in Jurkat cells cisplatin, HHT, mitoxantrone and HCPT 4 drug effects between the different dose groups at 2 h in Jurkat cells induced apoptosis was no significant difference in 4 hours and 8 hours between each group were significantly different (P <0.05, Figure 6). Drug-induced apoptosis in Jurkat cells followed by the intensity of MIT, HCPT, HHT, and CDDP.
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