解聚素-金属蛋白酶17-短发卡RNA转染骨髓间充质干细胞对人乳腺癌MCF-7细胞增殖的影响  

Effect of a disintegrin and metalloproteinase 17 short hairpin RNA transfection of bone marrow mesenchymal stem cells on proliferation of human breast cancer MCF-7 cells

作　　者：李长仔 胡宝山 吕捷 焦桂梅 张雪鹏 朱红成 Li Changzai;Hu Baoshan;Lyu Jie;Jiao Guimei;Zhang Xuepeng;Zhu Hongcheng(Department of Oncological Surgery,North China University of Science and Technology Affiliated Hospital,Tangshan 063000,China)

机构地区：[1]华北理工大学附属医院肿瘤外科,河北省唐山市063000

出　　处：《中国综合临床》2020年第5期440-444,共5页Clinical Medicine of China

基　　金：河北省卫计委医学科学研究项目(20191157)。

摘　　要：目的探讨解聚素-金属蛋白酶17(a disintegrin and metalloteinase 17,ADAM17)-短发卡RNA(short hairpin RNA,shRNA)转染骨髓间充质干细胞(bone marrow mesenchymal stem cell,BMSC)对人乳腺癌MCF-7细胞增殖的作用。方法设计4个针对ADAM17以重组慢病毒为载体的shRNA序列(ADAM17-hsa-297,ADAM17-hsa-1508,ADAM17-hsa-1658,ADAM17-hsa-1864)及1个阴性对照(LV10-NC),应用qPCR的方法筛选抑制率最佳的ADAM17-shRNA,实验分为3个组,即转染组、无意义序列组和对照组,按常规步骤提取RNA,进行逆转录以及扩增。用qPCR方法检测混合培养后ADAM17mRNA基因的表达,用MMT法测定人乳腺癌MCF-7细胞的增殖能力并进行统计学分析。结果对照组、无意义序列组和转染组24 h的吸光度值分别是0.270±0.040、0.250±0.035和0.185±0.080,组间比较差异有统计学意义(F=3.854,P=0.045),对照组与无意义序列组比较差异无统计学意义(P>0.05),对照组与转染组比较差异有统计学意义(P<0.05),无意义序列组与转染组差异无统计学意义(P>0.05);对照组、无意义序列组和转染组48 h的吸光度值分别是0.500±0.057、0.494±0.086和0.311±0.007,组间比较差异有统计学意义(F=19.42,P<0.001),转染组与对照组、无意义序列组比较差异均有统计学意义(P均<0.05),对照组与无意义序列组比较差异无统计学意义(P>0.05);对照组、无意义序列组和转染组72 h的吸光度值分别是0.720±0.150、0.713±0.174和0.558±0.071,组间比较差异无统计学意义(F=2.61,P=0.106)。结论ADAM17-shRNA转染的BMSC可以抑制24 h和48 h人乳腺癌MCF-7细胞的增殖,同时72 h人乳腺癌MCF-7细胞的增殖能力有下降趋势。Objective To investigate the effect of a disintegrin and metalloproteinase 17(ADAM17)-short hairpin RNA(shRNA)transfection on the proliferation of human breast cancer MCF-7 cells.Methods Four shRNA sequences(adam17-hsa-297,adam17-hsa-1508,adam17-hsa-1658)targeting ADAM17 were designed,Adam17-hsa-1864)and a negative control(LV10-NC)were used to screen the best inhibition rate of ADAM17 shRNA by qPCR.The experiment was divided into three groups:transfection group,meaningless sequence group and control group.RNA was extracted according to routine steps,and then reverse transcripted and amplified.The expression of ADAM17 mRNA was detected by qPCR and the proliferation of MCF-7 cells was measured by MMT method.Results The results of MTT assay showed that the absorbance values of control group,nonsense sequence group and transfection group were 0.270±0.040,0.250±0.035 and 0.185±0.080,respectively.There was significant difference between the two groups(F=3.854,P=0.045).There was no significant difference between the control group and the nonsignificant sequence group(P>0.05),and the difference between the control group and the transfection group was statistically significant(P<0.05).There was no significant difference between the nonsignificant sequence group and the transfection group(P>0.05);the absorbance values of the control group,the meaningless sequence group and the transfection group at 48 h were 0.500±0.057,0.494±0.086 and 0.311±0.007,respectively,and the differences between the two groups were statistically significant(F=19.42,P<0.001).There were significant differences between transfection group and control group and no significant sequence group(all P<0.05),but there was no significant difference between control group and non significant sequence group(P>0.05).The absorbance values of control group,nonsense sequence group and transfection group at 72 h were 0.720±0.150,0.713±0.174 and 0.558±0.071,respectively.There was no Conclusion BMSC transfected with ADAM17 shRNA could inhibit the proliferation

关 键 词：乳腺癌 解聚素-金属蛋白酶17 骨髓间充质干细胞 

分 类 号：R737.9[医药卫生—肿瘤]