title: Icaritin induces mitochondrial apoptosis by up-regulating miR-124 in human oral squamous cell carcinoma cells.
abstract: OBJECTIVE
  The present study is aimed to investigate the apoptosis-inducing effect of icaritin in human oral squamous cell carcinoma (OSCC) cells and the associated mechanisms.
  
  
  METHODS
  KB and SCC9 cell lines were used as model cell lines. Effect of icaritin on apoptosis was analyzed by flow cytometry. The effect of icaritin on mitochondrial apoptotic pathway was demonstrated by loss of mitochondrial membrane potential and release of cytocrome C from mitochondria. MiR-124 mimic and miR-124 inhibitor were used to manipulate the expression of miR-124 in OSCC cells. SiRNA targeting Sp1 and DNMT1 as well as Sp1 and DNMT1 overexpressing vector were utilized to confirm their roles in the apoptosis-inducing effect of icaritin in OSCC cells. Activation of relevant signaling pathway by icaritin and effect of icaritin on expression of targeting molecules were determined by western blots or qRT-PCR.
  
  
  RESULTS
  Our results showed that icaritin inhibited tumor cell viability in a dose- and time-dependent manner, and induced cell apoptosis via intrinsic mitochondrial pathway by upregulating miR-124. Moreover, our results showed that the icaritin exerted regulatory effect on miR-124 through suppressing Sp1/DNMT1 signaling.
  
  
  CONCLUSIONS
  Our data provide the first experimental evidence that icaritin induces mitochondrial apoptosis in OSCC cells by upregulating miR-124 and suggest a new mechanism to explain its anti-tumor effects.
authors.affiliation: Department of Oral & Maxillofacial Surgery, The First Affiliated Hospital, Zhengzhou University, China.
authors.forename: Limin
authors.initials: L
authors.lastname: Jin
authors.affiliation: Department of Nursing Management,The First Affiliated Hospital, Zhengzhou University, China.
authors.forename: Jinhong
authors.initials: J
authors.lastname: Miao
authors.affiliation: Department of Nursing Management,The First Affiliated Hospital, Zhengzhou University, China.
authors.forename: Yanjin
authors.initials: Y
authors.lastname: Liu
authors.affiliation: Department of Oral & Maxillofacial Surgery, The First Affiliated Hospital, Zhengzhou University, China.
authors.forename: Xingdan
authors.initials: X
authors.lastname: Li
authors.affiliation: Department of Oral & Maxillofacial Surgery, The First Affiliated Hospital, Zhengzhou University, China.
authors.forename: Yaqiong
authors.initials: Y
authors.lastname: Jie
authors.affiliation: Department of Oral & Maxillofacial Surgery, Stomatological Hospital of Nanyang, China.
authors.forename: Qianyun
authors.initials: Q
authors.lastname: Niu
authors.affiliation: Department of Oral & Maxillofacial Surgery, The First Affiliated Hospital, Zhengzhou University, China. Electronic address: drxingguanghan@163.com.
authors.forename: Xinguang
authors.initials: X
authors.lastname: Han
mesh_terms: D000970:Antineoplastic Agents / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects*; D002294:Carcinoma, Squamous Cell / Q000188:drug therapy* / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects; D045304:Cytochromes c / Q000378:metabolism; D000076123:DNA (Cytosine-5-)-Methyltransferase 1; D004248:DNA (Cytosine-5-)-Methyltransferases / Q000235:genetics / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D005419:Flavonoids / Q000494:pharmacology*; D015972:Gene Expression Regulation, Neoplastic; D006258:Head and Neck Neoplasms / Q000188:drug therapy* / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D006801:Humans; D053078:Membrane Potential, Mitochondrial / Q000187:drug effects; D035683:MicroRNAs / Q000235:genetics / Q000378:metabolism*; D008928:Mitochondria / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D009062:Mouth Neoplasms / Q000188:drug therapy* / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D034622:RNA Interference; D015398:Signal Transduction / Q000187:drug effects; D016329:Sp1 Transcription Factor / Q000235:genetics / Q000378:metabolism; D000077195:Squamous Cell Carcinoma of Head and Neck; D013997:Time Factors; D014162:Transfection; D015854:Up-Regulation
chemical_list: D000970:Antineoplastic Agents; D005419:Flavonoids; C509207:MIRN124 microRNA, human; D035683:MicroRNAs; D016329:Sp1 Transcription Factor; C108209:SP1 protein, human; D045304:Cytochromes c; D000076123:DNA (Cytosine-5-)-Methyltransferase 1; D004248:DNA (Cytosine-5-)-Methyltransferases; C000619380:DNMT1 protein, human; C499403:icaritin
keywords: Apoptosis; DNMT1; Icaritin; MiR-124; Sp1