基于JAK2/STAT3信号通路探讨青蒿琥酯对淋巴瘤Raji细胞生长及凋亡的影响

Analysis on the Effect of Artesunate on Growth and Apoptosis of Lymphoma Raji Cells Based on JAK2/STAT3 Signal Pathway

目的:基于Janus蛋白酪氨酸激酶2(JAK2)/信号转导和转录激活因子3(STAT3)信号通路探究青蒿琥酯(Art)对淋巴瘤Raji细胞生长及凋亡的影响,并探讨其可能的机制。方法:取对数期淋巴瘤Raji细胞,随机分为对照组、Art组、Colivelin组、Art+Colivelin组。对照组细胞常规培养,Art组细胞培养基内加入Art(400μg/mL),Colivelin组细胞培养基内加入Colivelin(0.5μmol/L),Art+Colivelin组细胞培养基内加入Art(400μg/mL)和Colivelin(0.5μmol/L)。MTT法检测细胞增殖能力;双染法检测细胞凋亡率;实时荧光定量聚合酶链式反应(qRT-PCR)法检测细胞肿瘤坏死因子-α(TNF-α)mRNA、白细胞介素-1β(IL-1β)mRNA相对表达量;免疫印迹法检测细胞pJAK2、JAK2、p-STAT3、STAT3蛋白相对表达量。结果:Art组细胞24、48、72h吸光度值,TNF-αmRNA、IL-1βmRNA相对表达量,以及JAK2/p-JAK2、STAT3/p-STAT3均低于对照组(P<0.05);Art组细胞凋亡率高于对照组(P<0.05);Colivelin组细胞24、48、72 h吸光度值,TNF-αmRNA、IL-1βmRNA相对表达量,以及JAK2/p-JAK2、STAT3/p-STAT3均高于对照组(P<0.05);Colivelin组细胞凋亡率低于对照组(P<0.05);Art+Colivelin组细胞24、48、72h吸光度值,TNF-αmRNA、IL-1βmRNA相对表达量,以及JAK2/p-JAK2、STAT3/p-STAT3均高于Art组(P<0.05);Art+Colivelin组细胞凋亡率低于Art组(P<0.05);Art+Colivelin组24、48、72 h吸光度值,TNF-αmRNA、IL-1βmRNA相对表达量,以及JAK2/p-JAK2、STAT3/p-STAT3均低于Colivelin组(P<0.05);Art+Colivelin组细胞凋亡率高于Colivelin组(P<0.05)。结论:Art可抑制淋巴瘤Raji细胞增殖及炎症反应,并能促进其凋亡,作用机制可能与抑制JAK2/STAT3信号通路有关。

Objective:To investigate the effect of artesunate(Art)on the growth and apoptosis of lymphoma Raji cells based on JAK2/STAT3 signal pathway,and to explore its possible mechanism.Methods:Raji cells from logarithmic phase lymphoma were randomly divided into control group,Art group,Colivelin group and Art+Colivelin group.The cells in the control group were routinely cultured.Art(400μg/mL)was added to the cell culture medium in Art group,Colivelin(0.5μmol/L)was added to the cell culture medium in Colivelin group,and the Art+Colivelin group was added with Art(400μg/mL)and Colivelin(0.5μmol/L).Cell proliferation was detected by MTT assay.Double staining was used to detect the apoptosis rate.The mRNA expressions of TNF-αand IL-1βwere detected by qRT-PCR.Western blot was used to detect the expression of p-JAK2,JAK2,p-STAT3 and STAT3 proteins in cells.Results:The absorbance values at 24,48 and 72 h,the relative expressions of TNF-αmRNA and IL-1βmRNA,and JAK2/p-JAK2,STAT3/p-STAT3 in Art group were lower than those in the control group(P<0.05).The apoptosis rate in Art group was higher than that in control group(P<0.05).The absorbance value at 24,48 and 72 h,the relative expression of TNF-αmRNA,IL-1βmRNA,and JAK2/p-JAK2,STAT3/p-STAT3 of cells in the Colivelin group were higher than those in the control group(P<0.05).The apoptosis rate in Colivelin group was lower than that in control group(P<0.05).The absorbance value at 24,48 and 72 h,the relative expression of TNF-αmRNA,IL-1βmRNA,and JAK2/p-JAK2,STAT3/p-STAT3 of cells in Art+Colivelin group were higher than those in Art group.The apoptosis rate in Art+Colivelin group was lower than that in Art group(P<0.05).The absorbance value at 24,48 and 72 h,the relative expression of TNF-αmRNA,IL-1βmRNA,and JAK2/p-JAK2,STAT3/p-STAT3 of cells in Art+Colivelin group were lower than those in Colivelin group.The apoptosis rate in Art+Colivelin group was higher than that in Colivelin group(P<0.05).Conclusion:Art can inhibit the proliferation and inflammatory response of lymphoma Raji cells,and promote their apoptosis.The mechanism of action may be related to the inhibition of JAK2/STAT3 signal pathway.