Study on the aging of leukemia K562 cells induced by ginsenoside Rg1 / 医学研究生学报

ABSTRACT

Objective　At present, the main studies of ginsenoside Rg1 are almost on the field of solid tumors and acute leukemias, and few on chronic leukemias. We aims to figure out the role of ATR-Chk1 pathway on cell aging in ginsenoside Rg1-treated leukemia K562 cells.　Methods　K562 cells were treated with ginsenoside Rg1 at different concentrations and divided into a control group (with 50 μL PBS culture solution) and 5 μmol/L ginsenoside group, 10 μmol/L ginsenoside group, 20 μmol/L ginsenoside group, 40 μmol/L ginsenoside group, 80 μmol/L ginsenoside group. CCK-8 assay，colony formation assay and flow cytometry for cell cycle detection were used to determine the effect of ginsenoside Rg1 on the aging of K562 cells. SA-β-Gal staining and Wright’s staining were used to observe the morphological changes of K562 cells’ aging. Real-time quantitative PCR and Western blot were used to detect the changes of ATR and Chk1 expression.　Results　The colony formation rate of K562 cells in the 20 μmol/L ginsenoside group was significantly lower than that in the other groups (P<0.05). CCK-8 test results showed that K562 cell proliferation of ginsenoside Rg1 induced groups was higher than that of the control group at 24, 48, and 72 hours (P<0.05). K562 cell proliferation inhibition rate was the highest in 20 μmol/L ginsenoside group for 48 hours treatment (P<0.05). The rate of SA-β-Gal positive cells [(95.833 ± 1.528) %] in 20 μmol/L ginsenoside-treated K562 cells for 48 h was significantly higher than that of the control group [(3.083 ± 0.764) %]. Cells blocked in G0/G1 phase and entered S and G2/M phases were significantly higher and lower than those in the control group, respectively (P<0.05).The ATR and Chk1 mRNA expression levels [(0.0117 ± 0.0038) %, (0.0120 ± 0.0021) %] were significantly higher than that of the control group ([0.0027 ± 0.0006) %, (0.0058 ± 0.0019) %) (P<0.05). ATR and Chk1 relative protein expression levels [(19.370 ± 0.994) %, (43.520 ± 1.236) %] were significantly increased compared with that of the control group [(17.080 ± 1.274) %, (39.100 ± 0.969) %) (P<0.05).　Conclusion　Ginsenoside Rg1 can induce the aging of K562 cells by regulating the ATR-Chk1 pathway, providing a new target for clinical leukemia treatment.