苦参碱联合化疗药物对人急性髓系白血病HL-60细胞增殖及侵袭力的影响(1)
摘要 目的：探究苦參碱联合化疗药物对人急性髓系白血病HL-60细胞增殖及侵袭力的影响。方法：体外培养人急性髓系白血病HL-60细胞，苦参碱单药组以1.0 g/L苦参碱干预，三氧化二砷单药组以3 μmol/L三氧化二砷处理，联合组以终浓度为1.0 g/L的苦参碱+3 μmol/L三氧化二砷处理，空白组以等量生理盐水处理。MTT法检测HL-60细胞增殖情况，ranswell小室实验检测HL-60侵袭力，蛋白免疫印迹法检测HL-60细胞MMPs蛋白表达情况。结果：苦参碱单药组、三氧化二砷单药组、联合组24 h PIR分别为(18.54±2.54)%，(29.54±2.62)%，(41.45±3.72)%，48 h PIR分别为(27.46±2.55)%，(38.93±3.76)%，(45.64±3.54)%，3组内HL-60的PIR均随着培养时间的增加而增加，其中联合组24 h、48 h PIR均高于同时间点的苦参碱单药组、三氧化二砷单药组，差异有统计学意义(P<0.05)。空白组、苦参碱单药组、三氧化二砷单药组、联合组侵袭细胞分别为(127.64±23.93)个、(94.24±15.23)个、(81.23±12.21)个、(75.13±10.69)个，相对侵袭指数分别为(100.00±0.00)%、(73.44±0.34)%、(62.47±0.15)%、(58.54±0.36)%;MMP-2蛋白相对表达量分别为(0.91±0.05)、(0.39±0.04)、(0.23±0.04)、(0.14±0.03);MMP-9蛋白相对表达量分别为(0.62±0.05)、(0.31±0.05)、(0.27±0.04)、(0.16±0.02)。与空白组比较，苦参碱单药组、三氧化二砷单药组、联合组侵袭细胞数量显著减少，相对侵袭指数及细胞中MMP-2、MMP-9相对表达量降低，且联合组均低于苦参碱单药组、三氧化二砷单药组，差异有统计学意义(P<0.05)。结论：苦参碱联合三氧化二砷通过下调MMP-2、MMP-9蛋白表达有效抑制人急性髓系白血病HL-60细胞增殖及侵袭。
关键词 人急性髓系白血病;HL-60细胞;凋亡;苦参碱;表达;机制
Effects of Matrine Combined with Chemotherapeutic Drugs on Proliferation and Invasion of Human Acute Myeloid Leukemia HL-60 Cells
Chao Rong，Hu Xiaoyan，Zhu Shengdong，Deng Wei，Wang Li
Abstract Objective：To investigate the effects of matrine combined with chemotherapeutic drugs on the proliferation and invasion of human acute myeloid leukemia HL-60 cells.Methods：Human acute myeloid leukemia HL-60 cells were cultured in vitro.Matrine monotherapy group was treated with 1.0 g/L matrine，arsenic trioxide monotherapy group with 3 mmol/L arsenic trioxide，combination group with 1.0 g/L matrine+3 mmol/L arsenic trioxide，blank group with the same amount of saline.MTT assay was used to detect the proliferation of HL-60 cells，ranswell chamber assay was used to detect the invasion ability of HL-60 cells，and Western blotting was used to detect the expression of MMPs in HL-60 cells.Results：The 24-hour PIR of matrine group，arsenic trioxide group and combination group were(18.54±2.54)%，(29.54±2.62)%，(41.45±3.72)% and 48-hour PIR were(27.46±2.55)%，(38.93±3.76)%，(45.64±3.54)%.The PIR of HL-60 in the 3 groups increased with the increase of culture time.PIR of 24 h and 48 h in THE combination group was higher than that of matrine and arsenic trioxide at the same time point(P<0.05).The number of invasive cells in the blank group，the matrine group，the arsenic trioxide group and the combination group were(127.64±23.93)，(94.24±15.23)，(81.23±12.21)，(75.13±10.69)，and the relative invasion index were(100.00±0.00)，(73.44±0.34)，(62.47±0.15)，(58.54±0.36)%，respectively.The relative expressions of MMP-2 were (0.91±0.05)，(0.39±0.04)，(0.23±0.04)，(0.14±0.03)，and the MMP-9 protein were (0.62±0.05)，(0.31±0.05)，(0.27±0.04)，(0.16±0.02)，respectively.Compared with the blank group，the number of invasive cells in matrine monotherapy group，arsenic trioxide monotherapy group and combination group decreased significantly，the relative invasion index and the relative expression of MMP-2 and MMP-9 protein in cells decreased，and the combined group was lower than matrine monotherapy group and arsenic trioxide monotherapy group(P<0.05).Conclusion：Matrine combined with arsenic trioxide can effectively inhibit the proliferation and invasion of human acute myeloid leukemia HL-60 cells，and its mechanism may be related to the down-regulation of MMP-2 and MMP-9 protein expression., 百拇医药(晁荣 胡晓燕 朱生东 邓伟 王莉)
关键词 人急性髓系白血病;HL-60细胞;凋亡;苦参碱;表达;机制
Effects of Matrine Combined with Chemotherapeutic Drugs on Proliferation and Invasion of Human Acute Myeloid Leukemia HL-60 Cells
Chao Rong，Hu Xiaoyan，Zhu Shengdong，Deng Wei，Wang Li
Abstract Objective：To investigate the effects of matrine combined with chemotherapeutic drugs on the proliferation and invasion of human acute myeloid leukemia HL-60 cells.Methods：Human acute myeloid leukemia HL-60 cells were cultured in vitro.Matrine monotherapy group was treated with 1.0 g/L matrine，arsenic trioxide monotherapy group with 3 mmol/L arsenic trioxide，combination group with 1.0 g/L matrine+3 mmol/L arsenic trioxide，blank group with the same amount of saline.MTT assay was used to detect the proliferation of HL-60 cells，ranswell chamber assay was used to detect the invasion ability of HL-60 cells，and Western blotting was used to detect the expression of MMPs in HL-60 cells.Results：The 24-hour PIR of matrine group，arsenic trioxide group and combination group were(18.54±2.54)%，(29.54±2.62)%，(41.45±3.72)% and 48-hour PIR were(27.46±2.55)%，(38.93±3.76)%，(45.64±3.54)%.The PIR of HL-60 in the 3 groups increased with the increase of culture time.PIR of 24 h and 48 h in THE combination group was higher than that of matrine and arsenic trioxide at the same time point(P<0.05).The number of invasive cells in the blank group，the matrine group，the arsenic trioxide group and the combination group were(127.64±23.93)，(94.24±15.23)，(81.23±12.21)，(75.13±10.69)，and the relative invasion index were(100.00±0.00)，(73.44±0.34)，(62.47±0.15)，(58.54±0.36)%，respectively.The relative expressions of MMP-2 were (0.91±0.05)，(0.39±0.04)，(0.23±0.04)，(0.14±0.03)，and the MMP-9 protein were (0.62±0.05)，(0.31±0.05)，(0.27±0.04)，(0.16±0.02)，respectively.Compared with the blank group，the number of invasive cells in matrine monotherapy group，arsenic trioxide monotherapy group and combination group decreased significantly，the relative invasion index and the relative expression of MMP-2 and MMP-9 protein in cells decreased，and the combined group was lower than matrine monotherapy group and arsenic trioxide monotherapy group(P<0.05).Conclusion：Matrine combined with arsenic trioxide can effectively inhibit the proliferation and invasion of human acute myeloid leukemia HL-60 cells，and its mechanism may be related to the down-regulation of MMP-2 and MMP-9 protein expression., 百拇医药(晁荣 胡晓燕 朱生东 邓伟 王莉)
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