title: Cisplatin selects for multidrug-resistant CD133+ cells in lung adenocarcinoma by activating Notch signaling.
abstract: Platinum-based chemotherapy is the first-line treatment for non-small cell lung cancer, but recurrence occurs in most patients. Recent evidence suggests that CD133(+) cells are the cause of drug resistance and tumor recurrence. However, the correlation between chemotherapy and regulation of CD133(+) cells has not been investigated methodically. In this study, we revealed that CD133(+) lung cancer cells labeled by a human CD133 promoter-driven GFP reporter exhibited drug resistance and stem cell characteristics. Treatment of H460 and H661 cell lines with low-dose cisplatin (IC(20)) was sufficient to enrich CD133(+) cells, to induce DNA damage responses, and to upregulate ABCG2 and ABCB1 expression, which therefore increased the cross-resistance to doxorubicin and paclitaxel. This cisplatin-induced enrichment of CD133(+) cells was mediated through Notch signaling as judged by increased levels of cleaved Notch1 (NICD1). Pretreatment with the γ-secretase inhibitor, N-[N-(3,5-difluorophenacetyl)-1-alanyl]-S-phenylglycine t-butyl ester (DAPT), or Notch1 short hairpin RNAs (shRNA) remarkably reduced the cisplatin-induced enrichment of CD133(+) cells and increased the sensitivity to doxorubicin and paclitaxel. Ectopic expression of NICD1 reversed the action of DAPT on drug sensitivity. Immunohistochemistry showed that CD133(+) cells were significantly increased in the relapsed tumors in three of six patients with lung cancer who have received cisplatin treatment. A similar effect was observed in animal experiments as cisplatin treatment increased Notch1 cleavage and the ratio of CD133(+) cells in engrafted tumors. Intratumoral injection of DAPT with cisplatin treatment significantly reduced CD133(+) cell number. Together, our results showed that cisplatin induces the enrichment of CD133(+) cells, leading to multidrug resistance by the activation of Notch signaling.
authors.affiliation: Institute of Clinical Medicine, National Cheng Kung University, Medical College, Tainan, Taiwan.
authors.forename: Yu-Peng
authors.initials: YP
authors.lastname: Liu
authors.forename: Chih-Jen
authors.initials: CJ
authors.lastname: Yang
authors.forename: Ming-Shyan
authors.initials: MS
authors.lastname: Huang
authors.forename: Chi-Tai
authors.initials: CT
authors.lastname: Yeh
authors.forename: Alexander T H
authors.initials: AT
authors.lastname: Wu
authors.forename: Yu-Cheng
authors.initials: YC
authors.lastname: Lee
authors.forename: Tsung-Ching
authors.initials: TC
authors.lastname: Lai
authors.forename: Chien-Hsin
authors.initials: CH
authors.lastname: Lee
authors.forename: Ya-Wen
authors.initials: YW
authors.lastname: Hsiao
authors.forename: Jean
authors.initials: J
authors.lastname: Lu
authors.forename: Chia-Ning
authors.initials: CN
authors.lastname: Shen
authors.forename: Pei-Jung
authors.initials: PJ
authors.lastname: Lu
authors.forename: Michael
authors.initials: M
authors.lastname: Hsiao
mesh_terms: D000071916:AC133 Antigen; D000230:Adenocarcinoma / Q000378:metabolism / Q000473:pathology*; D000077192:Adenocarcinoma of Lung; D000818:Animals; D015703:Antigens, CD / Q000096:biosynthesis*; D000970:Antineoplastic Agents / Q000494:pharmacology; D045744:Cell Line, Tumor; D002945:Cisplatin / Q000494:pharmacology; D018432:Drug Resistance, Multiple / Q000187:drug effects*; D019008:Drug Resistance, Neoplasm / Q000187:drug effects*; D006023:Glycoproteins / Q000096:biosynthesis*; D006801:Humans; D008175:Lung Neoplasms / Q000378:metabolism / Q000473:pathology*; D014411:Neoplastic Stem Cells / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D010455:Peptides; D051880:Receptors, Notch / Q000187:drug effects*; D015398:Signal Transduction / Q000502:physiology; D014183:Transplantation, Heterologous
chemical_list: D000071916:AC133 Antigen; D015703:Antigens, CD; D000970:Antineoplastic Agents; D006023:Glycoproteins; C000606023:PROM1 protein, human; D010455:Peptides; D051880:Receptors, Notch; D002945:Cisplatin