Standardized Polyalthia longifolia leaf extract induces the apoptotic HeLa cells death via microRNA regulation: identification, validation, and therapeutic potential

Polyalthia longifolia var. angustifolia Thw. (Annonaceae), is a famous traditional medicinal plant in Asia. Ample data specifies that the medicinal plant P. longifolia has anticancer activity; however, the detailed mechanisms of action still need to be well studied. Recent studies have revealed the cytotoxicity potential of P. longifolia leaf against HeLa cells. Therefore, the current study was conducted to examine the regulation of miRNAs in HeLa cancer cells treated with the standardized P. longifolia methanolic leaf extract (PLME). The regulation of miRNAs in HeLa cancer cells treated with the standardized PLME extract was studied through Illumina, Hi-Seq. 2000 platform of Next-Generation Sequencing (NGS) and various in silico bioinformatics tools. The PLME treatment regulated a subset of miRNAs in HeLa cells. Interestingly, the PLME treatment against HeLa cancer cells identified 10 upregulated and 43 downregulated (p < 0.05) miRNAs associated with apoptosis induction. Gene ontology (GO) term analysis indicated that PLME induces cell death in HeLa cells by inducing the pro-apoptotic genes. Moreover, the downregulated oncomiRs modulated by PLME treatment in HeLa cells were identified, targeting apoptosis-related genes through gene ontology and pathway analysis. The LC–ESI–MS/MS analysis identified the presence of Vidarabine and Anandamide compounds that were previously reported to exhibit anticancer activity. The findings of this study obviously linked the cell cytotoxicity effect of PLME treatment against the HeLa cells with regulating various miRNAs expression related to apoptosis induction in the HeLa cells. PLME treatment induced apoptotic HeLa cell death mechanism by regulating multiple miRNAs. The identified miRNAs regulated by PLME may provide further insight into the mechanisms that play a critical role in cervical cancer, as well as novel ideas regarding gene therapeutic strategies.

[1]  S. Sasidharan,et al.  Mechanisms of the In Vivo Antitumor Activity of Polyalthia longifolia Leaf Extract Against HeLa Cell Xenograft Tumor: A Microscopic-Based Histological and Immunohistochemical Microanalyses. , 2023, Microscopy and microanalysis : the official journal of Microscopy Society of America, Microbeam Analysis Society, Microscopical Society of Canada.

[2]  Jacques Ferlay,et al.  Estimates of incidence and mortality of cervical cancer in 2018: a worldwide analysis , 2019, The Lancet. Global health.

[3]  B. Reininger,et al.  Fruit and Vegetable Intake is Inversely Associated with Cancer Risk in Mexican-Americans , 2019, Nutrition and cancer.

[4]  W. Willett,et al.  Fruit and vegetable consumption and breast cancer incidence: Repeated measures over 30 years of follow‐up , 2019, International journal of cancer.

[5]  Jagat R Kanwar,et al.  Standardized Polyalthia longifolia leaf extract (PLME) inhibits cell proliferation and promotes apoptosis: The anti-cancer study with various microscopy methods. , 2017, Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie.

[6]  J. Kanwar,et al.  Polyalthia longifolia Methanolic Leaf Extracts (PLME) induce apoptosis, cell cycle arrest and mitochondrial potential depolarization by possibly modulating the redox status in hela cells. , 2017, Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie.

[7]  A. Etheridge,et al.  A systematic study on dysregulated microRNAs in cervical cancer development , 2016, International journal of cancer.

[8]  J. Yakisich,et al.  Role of apoptosis-related miRNAs in resveratrol-induced breast cancer cell death , 2016, Cell Death & Disease.

[9]  Q. Wei,et al.  MicroRNA-34a induces a senescence-like change via the down-regulation of SIRT1 and up-regulation of p53 protein in human esophageal squamous cancer cells with a wild-type p53 gene background. , 2016, Cancer letters.

[10]  Daniel J. Gaffney,et al.  A survey of best practices for RNA-seq data analysis , 2016, Genome Biology.

[11]  Pingfang Yang,et al.  Genome-Wide Dissection of the MicroRNA Expression Profile in Rice Embryo during Early Stages of Seed Germination , 2015, PloS one.

[12]  B. Zhang,et al.  A Comprehensive MicroRNA Expression Profile Related to Hypoxia Adaptation in the Tibetan Pig , 2015, PloS one.

[13]  Yan Guo,et al.  Three-stage quality control strategies for DNA re-sequencing data , 2014, Briefings Bioinform..

[14]  J. Ferrante,et al.  Update on prevention and screening of cervical cancer. , 2014, World journal of clinical oncology.

[15]  G. Ye,et al.  Fucoxanthin induces apoptosis in human cervical cancer cell line HeLa via PI3K/Akt pathway , 2014, Tumor Biology.

[16]  Changjun Tan,et al.  MiR-182 is up-regulated and targeting Cebpa in hepatocellular carcinoma. , 2014, Chinese journal of cancer research = Chung-kuo yen cheng yen chiu.

[17]  P. Levine,et al.  Methods and matrices: approaches to identifying miRNAs for Nasopharyngeal carcinoma , 2014, Journal of Translational Medicine.

[18]  S. Fogli,et al.  Anticancer activity of anandamide in human cutaneous melanoma cells. , 2013, European journal of pharmacology.

[19]  L. Yanqin,et al.  Emodin induces apoptosis of human cervical cancer hela cells via intrinsic mitochondrial and extrinsic death receptor pathway , 2013, Cancer Cell International.

[20]  J. Kanwar,et al.  Evaluation of the Genotoxic Potential against H2O2-Radical-Mediated DNA Damage and Acute Oral Toxicity of Standardized Extract of Polyalthia longifolia Leaf , 2013, Evidence-based complementary and alternative medicine : eCAM.

[21]  Edward Y. Chen,et al.  Enrichr: interactive and collaborative HTML5 gene list enrichment analysis tool , 2013, BMC Bioinformatics.

[22]  Nicholas A. Bokulich,et al.  Quality-filtering vastly improves diversity estimates from Illumina amplicon sequencing , 2012, Nature Methods.

[23]  A. Aziz,et al.  Antioxidant Activity and Hepatoprotective Potential of Polyalthia longifolia and Cassia spectabilis Leaves against Paracetamol-Induced Liver Injury , 2012, Evidence-based complementary and alternative medicine : eCAM.

[24]  Yue-Qin Chen,et al.  Camptothecin Induces Apoptosis in Cancer Cells via MicroRNA-125b-Mediated Mitochondrial Pathways , 2012, Molecular Pharmacology.

[25]  M. Siahpush,et al.  Socioeconomic, Rural-Urban, and Racial Inequalities in US Cancer Mortality: Part I—All Cancers and Lung Cancer and Part II—Colorectal, Prostate, Breast, and Cervical Cancers , 2012, Journal of cancer epidemiology.

[26]  M Arbyn,et al.  Worldwide burden of cervical cancer in 2008. , 2011, Annals of oncology : official journal of the European Society for Medical Oncology.

[27]  A. Srivastava,et al.  Identification of the antioxidant principles of Polyalthia longifolia var. pendula using TEAC assay , 2011, Natural product research.

[28]  E. Sibille,et al.  Human Postmortem Brain Research in Mental Illness Syndromes , 2011 .

[29]  H. Yin,et al.  Curcumin promotes apoptosis in A549/DDP multidrug-resistant human lung adenocarcinoma cells through an miRNA signaling pathway. , 2010, Biochemical and biophysical research communications.

[30]  Davis J. McCarthy,et al.  edgeR: a Bioconductor package for differential expression analysis of digital gene expression data , 2009, Bioinform..

[31]  Martin Reczko,et al.  The database of experimentally supported targets: a functional update of TarBase , 2008, Nucleic Acids Res..

[32]  Robert A Holt,et al.  The new paradigm of flow cell sequencing. , 2008, Genome research.

[33]  V. Beral,et al.  Cervical cancer and hormonal contraceptives: collaborative reanalysis of individual data for 16 573 women with cervical cancer and 35 509 women without cervical cancer from 24 epidemiological studies , 2007, The Lancet.

[34]  M. Vračko,et al.  Anticancer Activity of Selected Phenolic Compounds: QSAR Studies Using Ridge Regression and Neural Networks , 2007, Chemical biology & drug design.

[35]  Martti T. Tammi,et al.  MicroTar: predicting microRNA targets from RNA duplexes , 2006, BMC Bioinformatics.

[36]  P. D. Karp,et al.  The outcomes of pathway database computations depend on pathway ontology , 2006, Nucleic acids research.

[37]  Jan Krüger,et al.  RNAhybrid: microRNA target prediction easy, fast and flexible , 2006, Nucleic Acids Res..

[38]  Walter Sneader,et al.  Drug Discovery (The History) , 2005 .

[39]  A. Al-Hazzani,et al.  Novel Marine Compounds: Anticancer or Genotoxic? , 2004, Journal of biomedicine & biotechnology.

[40]  R. Ruffolo,et al.  Drug discovery , 2005, Nature Biotechnology.

[41]  Xianzhong Wu,et al.  Antioxidant and antiproliferative activities of common fruits. , 2002, Journal of agricultural and food chemistry.

[42]  S. Jang,et al.  Nitric oxide as a pro-apoptotic as well as anti-apoptotic modulator. , 2002, Journal of biochemistry and molecular biology.

[43]  M. Ashburner,et al.  Gene Ontology: tool for the unification of biology , 2000, Nature Genetics.

[44]  Dean P. Jones,et al.  Mitochondrial control of apoptosis: the role of cytochrome c. , 1998, Biochimica et biophysica acta.

[45]  P Green,et al.  Base-calling of automated sequencer traces using phred. II. Error probabilities. , 1998, Genome research.

[46]  P. Green,et al.  Base-calling of automated sequencer traces using phred. I. Accuracy assessment. , 1998, Genome research.

[47]  R. Moore.  Handbook of molecular and cellular methods in biology and medicine , 1996 .

[48]  V. Ambros,et al.  The C. elegans heterochronic gene lin-4 encodes small RNAs with antisense complementarity to lin-14 , 1993, Cell.

[49]  H. Boedtker,et al.  RNA molecular weight determinations by gel electrophoresis under denaturing conditions, a critical reexamination. , 1977, Biochemistry.

[50]  K. Alexandrov,et al.  Fractionation of Ribonucleic Acids by ‘Sephadex’ Agarose Gel Electrophoresis , 1968, Nature.

[51]  C. Paoletti,et al.  A new fluorometric method for RNA and DNA determination. , 1966, Analytical biochemistry.

[52]  J. L. SIMONSEN,et al.  The Wealth of India , 1949, Nature.

[53]  Soonhag Kim,et al.  Theragnosis-based combined cancer therapy using doxorubicin-conjugated microRNA-221 molecular beacon. , 2016, Biomaterials.

[54]  A. Aziz,et al.  Radioprotective activity of Polyalthia longifolia standardized extract against X-ray radiation injury in mice. , 2016, Physica medica : PM : an international journal devoted to the applications of physics to medicine and biology : official journal of the Italian Association of Biomedical Physics.

[55]  Torgeir R. Hvidsten,et al.  Guidelines for RNA-Seq data analysis , 2014 .

[56]  A. Goyal,et al.  Comparative study of extraction, purification and estimation of bromelain from stem and fruit of pineapple plant , 2010, The Thai Journal of Pharmaceutical Sciences.

[57]  Brad T. Sherman,et al.  Nucleic Acids Research Advance Access published June 18, 2007 , 2007 .

[58]  S. Lightfoot,et al.  Quantitation comparison of total RNA using the Agilent , 2002 .

[59]  International Human Genome Sequencing Consortium.  Initial sequencing and analysis of the human genome , 2001, Nature.

[60]  Y. Benjamini,et al.  Controlling the false discovery rate: a practical and powerful approach to multiple testing , 1995 .

[61]  J. Glasel.  Validity of nucleic acid purities monitored by 260nm/280nm absorbance ratios. , 1995, BioTechniques.

[62]  C. A. Thomas,et al.  Molecular cloning. , 1977, Advances in pathobiology.