摘要
目的探讨CUEDC2与多发性骨髓瘤(MM)细胞糖皮质激素敏感性的关系。方法培养U266、RPMI8226细胞至铺满培养瓶80%以上时进行传代,取对数生长期U266、RPMI8226细胞,分别以1×10^5 L^-1的密度接种于96孔培养板,将U266、RPMI8226细胞分别分为对照组(未经药物处理)和地塞米松(DEX)组(DEX终质量浓度分别为50、75、100、150、200 mg·L^-1)、N-乙酰基-L-亮氨酰-L-亮氨酰-L-正亮氨酸(ALLN)组(ALLN终质量浓度为25、50、125、250 mg·L^-1)、ALLN+DEX组(25 mg·L^-1 ALLN+50 mg·L^-1 DEX),采用四甲基偶氮唑蓝法检测U266、RPMI8226细胞增殖抑制率,聚合酶链式反应法检测U266、RPMI8226细胞内CUEDC2 mRNA表达。结果DEX和ALLN对U266、RPMI8226细胞增殖均有抑制作用,随着DEX、ALLN质量浓度增加,抑制作用逐渐增强(P<0.05)。DEX对RPMI8226和U266细胞的半抑制浓度(IC 50)分别为(68.25±3.81)、(95.92±4.92)mg·L^-1,DEX对RPMI8226细胞的IC 50显著低于U266细胞(P<0.05)。ALLN对RPMI8226和U266细胞的IC 50分别为(122.50±4.46)、(112.70±11.03)mg·L^-1,ALLN对RPMI8226和U266细胞的IC 50比较差异无统计学意义(P>0.05)。ALLN+DEX组U266、RPMI8226细胞增殖抑制率高于DEX组(50 mg·L^-1)和ALLN组(25 mg·L^-1)(P<0.05),ALLN组(25 mg·L^-1)U266细胞增殖抑制率高于DEX组(50 mg·L^-1)(P<0.05),ALLN组(25 mg·L^-1)与DEX组(50 mg·L^-1)RPMI8226细胞增殖抑制率比较差异无统计学意义(P>0.05);DEX组(50 mg·L^-1)RPMI 8226细胞增殖抑制率高于U266细胞(P<0.05),ALLN组(25 mg·L^-1)、ALLN+DEX组RPMI 8226细胞与U266细胞增殖抑制率比较差异无统计学意义(P>0.05)。DEX组与对照组U266和RPMI8226细胞中CUEDC2 mRNA表达比较差异无统计学意义(P>0.05),ALLN组、ALLN+DEX组U266和RPMI8226细胞中CUEDC2 mRNA表达低于对照组和DEX组(P<0.05),ALLN+DEX组与ALLN组U266和RPMI8226细胞中CUEDC2 mRNA表达比较差异无统计学意义(P>0.05),对照组、DEX组U266细胞中CUEDC2 mRNA表达高于RPMI8226细胞(P<0.05),ALLN组、ALLN+DEX组U266细胞与RPMI8226细胞中CUEDC2 mRNA表达比较差异无统计学意义(P>0.05)。结论CUEDC2表达可能与MM细胞对糖皮质激素敏感性有关,高表达CUEDC2的MM细胞对DEX的敏感性较差,抑制CUEDC2表达可提高MM细胞对DEX的敏感性。
Objective To investigate the correlation between CUEDC2 and the glucocorticoid sensitivity in multiple myeloma(MM)cells.Methods The U266 and RPMI8226 cells were subcultured when they overspread more than 80%of the culture flask.The U266 and RPMI8226 cells in the logarithmic growth phase were respectively inoculated on the 96-well culture plate at the density of 1×10^5 L^-1.The U266 and RPMI8226 cells were divided into control group(without drug treatment),dexamethasone(DEX)group(the final mass concentration of DEX were 50,75,100,150 and 200 mg·L^-1),N-acetyl-leu-leu-norleucine(ALLN)group(the final mass concentration of ALLN were 25,50,125 and 250 mg·L^-1)and ALLN+DEX group(25 mg·L^-1 ALLN+50 mg·L^-1 DEX).The proliferation inhibition rate of U266 and RPMI8226 cells was detected by methyl thiazolyl tetrazolium(MTT)method.The expression of CUEDC2 mRNA in U266 and RPMI8226 cells was detected by polymerase chain reaction.Results Both DEX and ALLN had inhibitory effect on the proliferation of U266 and RPMI8226 cells,and the inhibitory effect was gradually enhanced with the increase of the mass concentration of DEX and ALLN(P<0.05).The 50%inhibiting concentration(IC 50)of DEX to the RPMI8226 and U266 cells was(68.25±3.81)and(95.92±4.92)mg·L^-1,respectively;the IC 50 of DEX to RPMI8226 cells was significantly lower than that to U266 cells(P<0.05).The IC 50 of ALLN to the RPMI8226 and U266 cells was(122.50±4.46)and(112.70±11.03)mg·L^-1,respectively;there was no significant difference in the IC 50 of ALLN between RPMI8226 and U266 cells(P>0.05).The proliferation inhibition rate of U266 and RPMI8226 cells in the ALLN+DEX group was higher than that in the DEX group(50 mg·L^-1)and the ALLN group(25 mg·L^-1)(P<0.05).The proliferation inhibition rate of U266 cells in the ALLN group(25 mg·L^-1)was higher than that in the DEX group(50 mg·L^-1)(P<0.05).There was no significant difference in the proliferation inhibition rate of RPMI8226 cells between the ALLN group(25 mg·L^-1)and the DEX group(50 mg·L^-1)(P>0.05).The proliferation inhibition rate of RPMI 8226 cells was higher than that of U266 cells in the DEX group(P<0.05).There was no significant difference in the proliferation inhibition rate of RPMI 8226 cells and U266 cells between the ALLN group(25 mg·L^-1)and the ALLN+DEX group(P>0.05).There was no significant difference in the expression of CUEDC2 mRNA in U266 and RPMI8226 cells between the DEX group and the control group(P>0.05).The expression of CUEDC2 mRNA in U266 and RPMI8226 cells of the ALLN group,ALLN+DEX group was lower than that of control group and DEX group(P<0.05).There was no significant difference in the expression of CUEDC2 mRNA between the ALLN+DEX group and the ALLN group(P>0.05).The expression of CUEDC2 mRNA in U266 cells was higher than that in RPMI8226 cells in the control group and DEX group(P<0.05).There was no significant difference in the expression of CUEDC2 mRNA between U266 cells and RPMI8226 cells in the ALLN group and ALLN+DEX group(P>0.05).Conclusion The expression of CUEDC2 may be related to the sensitivity of MM cells to glucocorticoid,the MM cells with high expression of CUEDC2 were less sensitive to DEX,inhibiting the CUEDC2 expression can improve the sensitivity of MM cells to DEX.
作者
杨翠
李敬东
王婉玲
字友梅
高攀科
韩效林
YANG Cui;LI Jing-dong;WANG Wan-ling;ZI You-mei;GAO Pan-ke;HAN Xiao-lin(Department of Hematology,the First Affiliated Hospital of Xinxiang Medical University,Weihui 453100,Henan Province,China)
出处
《新乡医学院学报》
CAS
2019年第12期1125-1129,共5页
Journal of Xinxiang Medical University
基金
河南省医学科技攻关项目(编号:201602152)
