title: The Hedgehog signalling pathway mediates drug response of MCF-7 mammosphere cells in breast cancer patients.
abstract: BCSCs (breast cancer stem cells) have been shown to be resistant to chemotherapy. However, the mechanisms underlying BCSC-mediated chemoresistance remain poorly understood. The Hh (Hedgehog) pathway is important in the stemness maintenance of CSCs. Nonetheless, it is unknown whether the Hh pathway is involved in BCSC-mediated chemoresistance. In the present study, we cultured breast cancer MCF-7 cells in suspension in serum-free medium to obtain BCSC-enriched MCF-7 MS (MCF-7 mammosphere) cells. We showed that MCF-7 MS cells are sensitive to salinomycin, but not paclitaxel, distinct from parent MCF-7 cells. The expression of the critical components of Hh pathway, i.e., PTCH (Patched), SMO (Smoothened), Gli1 and Gli2, was significantly up-regulated in MCF-7 MS cells; salinomycin, but not paclitaxel, treatment caused a remarkable decrease in expression of those genes in MCF-7 MS cells, but not in MCF-7 cells. Salinomycin, but not paclitaxel, increased apoptosis, decreased the migration capacity of MCF-7 MS cells, accompanied by a decreased expression of c-Myc, Bcl-2 and Snail, the target genes of the Hh pathway. The salinomycin-induced cytotoxic effect could be blocked by Shh (Sonic Hedgehog)-mediated Hh signalling activation. Inhibition of the Hh pathway by cyclopamine could sensitize MCF-7 MS cells to paclitaxel. In addition, salinomycin, but not paclitaxel, significantly reduced the tumour growth, accompanied by decreased expression of PTCH, SMO, Gli1 and Gli2 in xenograft tumours. Furthermore, the expression of SMO and Gli1 was positively correlated with the expression of CD44+ / CD24-, and the expression of SMO and Gli1 in CD44+ / CD24- tissues was associated with a significantly shorter OS (overall survival) and DFS (disease-free survival) in breast cancer patients receiving chemotherapy.
authors.affiliation: Department of Pharmacology, School of Pharmacy, China Medical University, Shenyang, Liaoning Province, China.
authors.forename: Miao
authors.initials: M
authors.lastname: He
authors.affiliation: Department of Pharmacology, School of Pharmacy, China Medical University, Shenyang, Liaoning Province, China.
authors.forename: Yingzi
authors.initials: Y
authors.lastname: Fu
authors.affiliation: Department of Pharmacology, School of Pharmacy, China Medical University, Shenyang, Liaoning Province, China.
authors.forename: Yuanyuan
authors.initials: Y
authors.lastname: Yan
authors.affiliation: Department of Pharmacology, School of Pharmacy, China Medical University, Shenyang, Liaoning Province, China.
authors.forename: Qinghuan
authors.initials: Q
authors.lastname: Xiao
authors.affiliation: Department of Pharmacology, School of Pharmacy, China Medical University, Shenyang, Liaoning Province, China.
authors.forename: Huizhe
authors.initials: H
authors.lastname: Wu
authors.affiliation: Department of Pharmacology, School of Pharmacy, China Medical University, Shenyang, Liaoning Province, China.
authors.forename: Weifan
authors.initials: W
authors.lastname: Yao
authors.affiliation: Department of Pharmacology, School of Pharmacy, China Medical University, Shenyang, Liaoning Province, China.
authors.forename: Haishan
authors.initials: H
authors.lastname: Zhao
authors.affiliation: Department of Pharmacology, School of Pharmacy, China Medical University, Shenyang, Liaoning Province, China.
authors.forename: Lin
authors.initials: L
authors.lastname: Zhao
authors.affiliation: Department of Pharmacology, School of Pharmacy, China Medical University, Shenyang, Liaoning Province, China.
authors.forename: Qian
authors.initials: Q
authors.lastname: Jiang
authors.affiliation: Department of Pharmacology, School of Pharmacy, China Medical University, Shenyang, Liaoning Province, China.
authors.forename: Zhaojin
authors.initials: Z
authors.lastname: Yu
authors.affiliation: Department of Surgical Oncology, The First Affiliated Hospital of China Medical University, Shenyang, Liaoning Province, China.
authors.forename: Feng
authors.initials: F
authors.lastname: Jin
authors.affiliation: Institute of Pathology and Pathophysiology, China Medical University, Shenyang, Liaoning Province, China.
authors.forename: Xiaoyi
authors.initials: X
authors.lastname: Mi
authors.affiliation: Institute of Pathology and Pathophysiology, China Medical University, Shenyang, Liaoning Province, China.
authors.forename: Enhua
authors.initials: E
authors.lastname: Wang
authors.affiliation: Center of the Laboratory Technology and Experimental Medicine, China Medical University, Shenyang, Liaoning Province, China.
authors.forename: Zeshi
authors.initials: Z
authors.lastname: Cui
authors.affiliation: State Key Laboratory of Oncology in South China, Cancer Center, Sun Yat-Sen University, Guangzhou, China.
authors.forename: Liwu
authors.initials: L
authors.lastname: Fu
authors.affiliation: Department of Medicine, University of Chicago, Chicago, IL 60637, U.S.A.
authors.forename: Jianju
authors.initials: J
authors.lastname: Chen
authors.affiliation: Department of Pharmacology, School of Pharmacy, China Medical University, Shenyang, Liaoning Province, China Institute of Pathology and Pathophysiology, China Medical University, Shenyang, Liaoning Province, China weiminjiecmu@163.com.
authors.forename: Minjie
authors.initials: M
authors.lastname: Wei
mesh_terms: D000818:Animals; D000970:Antineoplastic Agents / Q000494:pharmacology / Q000627:therapeutic use; D001943:Breast Neoplasms / Q000188:drug therapy / Q000235:genetics / Q000378:metabolism*; D051927:CD24 Antigen / Q000378:metabolism; D019008:Drug Resistance, Neoplasm / Q000187:drug effects / Q000235:genetics; D005260:Female; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects; D053823:Hedgehog Proteins / Q000378:metabolism*; D006801:Humans; D018960:Hyaluronan Receptors / Q000378:metabolism; D053208:Kaplan-Meier Estimate; D051741:Kruppel-Like Transcription Factors / Q000235:genetics / Q000378:metabolism; D061986:MCF-7 Cells; D008807:Mice, Inbred BALB C; D008819:Mice, Nude; D014411:Neoplastic Stem Cells / Q000187:drug effects / Q000378:metabolism*; D009687:Nuclear Proteins / Q000235:genetics / Q000378:metabolism; D017239:Paclitaxel / Q000494:pharmacology / Q000627:therapeutic use; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000235:genetics / Q000378:metabolism; D016271:Proto-Oncogene Proteins c-myc / Q000235:genetics / Q000378:metabolism; D011714:Pyrans / Q000494:pharmacology / Q000627:therapeutic use; D043562:Receptors, G-Protein-Coupled / Q000235:genetics / Q000378:metabolism; D015398:Signal Transduction; D000072086:Smoothened Receptor; D018874:Spheroids, Cellular / Q000187:drug effects / Q000378:metabolism; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism; D023041:Xenograft Model Antitumor Assays; D000071676:Zinc Finger Protein GLI1; D000075746:Zinc Finger Protein Gli2; D000095662:Polyether Polyketides
chemical_list: D000970:Antineoplastic Agents; D051927:CD24 Antigen; D053823:Hedgehog Proteins; D018960:Hyaluronan Receptors; D051741:Kruppel-Like Transcription Factors; D009687:Nuclear Proteins; D017239:Paclitaxel; D019253:Proto-Oncogene Proteins c-bcl-2; D016271:Proto-Oncogene Proteins c-myc; D011714:Pyrans; D043562:Receptors, G-Protein-Coupled; D000072086:Smoothened Receptor; D014157:Transcription Factors; D000071676:Zinc Finger Protein GLI1; D000075746:Zinc Finger Protein Gli2; C497483:CD44 protein, human; C400551:GLI1 protein, human; C082593:GLI2 protein, human; C489427:MYC protein, human; C491023:SMO protein, human; C010327:salinomycin; D000095662:Polyether Polyketides
keywords: Hedgehog signalling pathway; breast cancer tissues; chemoresistance; mammosphere