title: Mineral pitch induces apoptosis and inhibits proliferation via modulating reactive oxygen species in hepatic cancer cells.
abstract: BACKGROUND
  Mineral Pitch (MP) is a dark brown coloured humic matter originating from high altitude rocks. It is an Ayurvedic medicinal food, commonly used by the people of the Himalayan regions of Nepal and India for various body ailments.
  
  
  METHODS
  The Huh-7 cells were treated with different concentrations of MP for 24 h, and both apoptosis and proliferation was determined by the TUNEL and MTT assays respectively. The formation of ROS and nitric oxide was analysed by DCFH-DA and Griess reagent respectively. The expression of miRNA-21 and miRNA-22 were checked by the real time PCR. Effect of miRNA-22 on proliferation and c-myc was studied by over-expressing miRNA-22 premiRs in Huh-7 cells.
  
  
  RESULTS
  We found that MP enhanced anti-cancer effects by inducing apoptosis and inhibiting proliferation. MP induced both ROS and NO, upon neutralizing them, there was a partial recovery of apoptosis and proliferation. MP also induced miRNA-22 expression, while miRNA-21 expression was inhibited. Over-expression of miRNA-22 resulted in a significant inhibition of proliferation. miRNA-22 directly targeted c-myc gene, thereby inhibited proliferation. These results clearly show that MP induces its anti-cancer activity by more than one pathway.
  
  
  CONCLUSIONS
  The data clearly indicate that MP induced apoptosis via the production of ROS, and inhibited proliferation by inducing miRNA-22 and inhibiting miRNA-21 in Huh-7 cells.
authors.affiliation: Faculty of Life Sciences and Biotechnology, South Asian University, Akbar Bhawan Chanakyapuri, New Delhi, India. pant.kishor1@gmail.com.
authors.forename: Kishor
authors.initials: K
authors.lastname: Pant
authors.affiliation: Faculty of Life Sciences and Biotechnology, South Asian University, Akbar Bhawan Chanakyapuri, New Delhi, India.
authors.forename: Parul
authors.initials: P
authors.lastname: Gupta
authors.affiliation: Faculty of Life Sciences and Biotechnology, South Asian University, Akbar Bhawan Chanakyapuri, New Delhi, India.
authors.forename: Preeti
authors.initials: P
authors.lastname: Damania
authors.affiliation: Faculty of Life Sciences and Biotechnology, South Asian University, Akbar Bhawan Chanakyapuri, New Delhi, India.
authors.forename: Ajay K
authors.initials: AK
authors.lastname: Yadav
authors.affiliation: Faculty of Life Sciences and Biotechnology, South Asian University, Akbar Bhawan Chanakyapuri, New Delhi, India.
authors.forename: Aanchal
authors.initials: A
authors.lastname: Gupta
authors.affiliation: Faculty of Life Sciences and Biotechnology, South Asian University, Akbar Bhawan Chanakyapuri, New Delhi, India.
authors.forename: Anam
authors.initials: A
authors.lastname: Ashraf
authors.affiliation: Faculty of Life Sciences and Biotechnology, South Asian University, Akbar Bhawan Chanakyapuri, New Delhi, India. drsenthil@sau.ac.in.
authors.forename: Senthil K
authors.initials: SK
authors.lastname: Venugopal
mesh_terms: D000972:Antineoplastic Agents, Phytogenic / Q000627:therapeutic use*; D000975:Antioxidants / Q000378:metabolism; D017209:Apoptosis / Q000187:drug effects*; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D004354:Drug Screening Assays, Antitumor; D022781:Hepatocytes; D006801:Humans; D006812:Humic Substances; D008113:Liver Neoplasms / Q000188:drug therapy*; D008519:Medicine, Traditional; D035683:MicroRNAs / Q000378:metabolism; D008903:Minerals / Q000627:therapeutic use*; D017382:Reactive Oxygen Species / Q000378:metabolism; D012116:Resins, Plant / Q000627:therapeutic use*
chemical_list: D000972:Antineoplastic Agents, Phytogenic; D000975:Antioxidants; D006812:Humic Substances; C524940:MIRN21 microRNA, human; C548375:MIRN22 microRNA, human; D035683:MicroRNAs; D008903:Minerals; D017382:Reactive Oxygen Species; D012116:Resins, Plant; C045109:mumie
keywords: Anticancer; Hepatocellular carcinoma; Mineral Pitch; Oxidative stress; Proliferation