title: A small hairpin RNA targeting myeloid cell leukemia-1 enhances apoptosis in host macrophages infected with Mycobacterium tuberculosis.
abstract: Myeloid cell leukemia-1 (Mcl-1) plays an important role in various cell survival pathways. Some studies indicated that the expression of Mcl-1 was upregulated in host cells during infection with the virulent Mycobacterium tuberculosis strain, H37Rv. The present study was designed to investigate the effect of inhibiting Mcl-1 expression both in vivo and in vitro on apoptosis of host macrophages infected with M. tuberculosis using a small hairpin (sh)RNA. Mcl-1 expression was detected by the real time-polymerase chain reaction, western blotting, and immunohistochemistry. Flow cytometry and transmission electron microscopy were used to measure host macrophage apoptosis. We found elevated Mcl-1 levels in host macrophages infected with M. tuberculosis H37Rv. The expression of Mcl-1 was downregulated efficiently in H37Rv-infected host macrophages using shRNA. Knockdown of Mcl-1 enhanced the extent of apoptosis in H37Rv-infected host macrophages significantly. The increased apoptosis correlated with a decrease in M. tuberculosis colony forming units recovered from H37Rv-infected cells that were treated with Mcl-1-shRNA. Reducing Mcl-1 accumulation by shRNA also reduced accumulation of the anti-apoptotic gene, Bcl-2, and increased expression of the pro-apoptotic gene, Bax, in H37Rv-infected host macrophages. Our results showed that specific knockdown of Mcl-1 expression increased apoptosis of host macrophages significantly and decreased the intracellular survival of a virulent strain of M. tuberculosis. These data indicate that interference with Mcl-1 expression may provide a new avenue for tuberculosis therapy.
authors.affiliation: Department of Pathophysiology, Medical College of Shihezi University, Xinjiang, P. R. China.|Key Laboratory of Xinjiang Endemic and Ethnic Diseases Cooperated by Education Ministry with Xinjiang Province, Xinjiang, Shihezi, P. R. China.
authors.forename: Fei-yu
authors.initials: FY
authors.lastname: Wang
authors.affiliation: Department of Pathophysiology, Medical College of Shihezi University, Xinjiang, P. R. China.|Key Laboratory of Xinjiang Endemic and Ethnic Diseases Cooperated by Education Ministry with Xinjiang Province, Xinjiang, Shihezi, P. R. China.
authors.forename: Yu-qing
authors.initials: YQ
authors.lastname: Zhang
authors.affiliation: Department of Urinary Surgery, the First Affiliated Hospital, Xinjiang, Shihezi, P. R. China.|Key Laboratory of Xinjiang Endemic and Ethnic Diseases Cooperated by Education Ministry with Xinjiang Province, Xinjiang, Shihezi, P. R. China.
authors.forename: Xin-min
authors.initials: XM
authors.lastname: Wang
authors.affiliation: Department of Pathogen Biology and Immunology, Medical College of Shihezi University, Xinjiang, P. R. China.|Key Laboratory of Xinjiang Endemic and Ethnic Diseases Cooperated by Education Ministry with Xinjiang Province, Xinjiang, Shihezi, P. R. China.
authors.forename: Chan
authors.initials: C
authors.lastname: Wang
authors.affiliation: Department of Pathophysiology, Medical College of Shihezi University, Xinjiang, P. R. China.|Key Laboratory of Xinjiang Endemic and Ethnic Diseases Cooperated by Education Ministry with Xinjiang Province, Xinjiang, Shihezi, P. R. China.
authors.forename: Xiao-fang
authors.initials: XF
authors.lastname: Wang
authors.affiliation: Department of Pathophysiology, Medical College of Shihezi University, Xinjiang, P. R. China.|Key Laboratory of Xinjiang Endemic and Ethnic Diseases Cooperated by Education Ministry with Xinjiang Province, Xinjiang, Shihezi, P. R. China.
authors.forename: Jiang-dong
authors.initials: JD
authors.lastname: Wu
authors.affiliation: Department of Pathophysiology, Medical College of Shihezi University, Xinjiang, P. R. China.|Key Laboratory of Xinjiang Endemic and Ethnic Diseases Cooperated by Education Ministry with Xinjiang Province, Xinjiang, Shihezi, P. R. China.
authors.forename: Fang
authors.initials: F
authors.lastname: Wu
authors.affiliation: Department of Pathophysiology, Medical College of Shihezi University, Xinjiang, P. R. China.|Key Laboratory of Xinjiang Endemic and Ethnic Diseases Cooperated by Education Ministry with Xinjiang Province, Xinjiang, Shihezi, P. R. China.
authors.forename: Wan-jiang
authors.initials: WJ
authors.lastname: Zhang
authors.affiliation: Department of Pathophysiology, Medical College of Shihezi University, Xinjiang, P. R. China. 1257067540@qq.com.|Key Laboratory of Xinjiang Endemic and Ethnic Diseases Cooperated by Education Ministry with Xinjiang Province, Xinjiang, Shihezi, P. R. China. 1257067540@qq.com.
authors.forename: Le
authors.initials: L
authors.lastname: Zhang
mesh_terms: D000818:Animals; D017209:Apoptosis; D002467:Cell Nucleus / Q000378:metabolism; D003593:Cytoplasm / Q000378:metabolism; D005260:Female; D005786:Gene Expression Regulation; D008264:Macrophages / Q000166:cytology / Q000378:metabolism / Q000382:microbiology*; D017737:Macrophages, Peritoneal / Q000166:cytology / Q000378:metabolism* / Q000382:microbiology*; D051379:Mice; D008807:Mice, Inbred BALB C; D050296:Microbial Viability; D009169:Mycobacterium tuberculosis / Q000502:physiology*; D064549:Myeloid Cell Leukemia Sequence 1 Protein / Q000235:genetics* / Q000378:metabolism*; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000235:genetics / Q000378:metabolism; D000067996:RAW 264.7 Cells; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000235:genetics*; D015854:Up-Regulation; D051028:bcl-2-Associated X Protein / Q000235:genetics / Q000378:metabolism
chemical_list: C494827:Bax protein, mouse; C579094:Mcl1 protein, mouse; D064549:Myeloid Cell Leukemia Sequence 1 Protein; D019253:Proto-Oncogene Proteins c-bcl-2; D034741:RNA, Small Interfering; D051028:bcl-2-Associated X Protein; C052617:Bcl2 protein, mouse
keywords: Mcl-1; Mcl-1-shRNA; Mycobacterium tuberculosis; apoptosis; host macrophages