Characterization and phytochemical constituents of Periploca hydaspidis Falc crude extract and its anticancer activities

Saima Ali; Muhammad Rashid Khan; Riffat Batool; Sayed Afzal Shah; Javed Iqbal; Banzeer Ahsan Abbasi; Tabassum Yaseen; Nida Zahra; Adil Aldhahrani; Fayez Althobaiti

doi:10.1016/j.sjbs.2021.08.020

Characterization and phytochemical constituents of Periploca hydaspidis Falc crude extract and its anticancer activities

Saudi J Biol Sci. 2021 Oct;28(10):5500-5517. doi: 10.1016/j.sjbs.2021.08.020. Epub 2021 Aug 12.

Authors

Affiliations

¹ Department of Biochemistry, Faculty of Biological Sciences, Quaid-i-Azam University, Islamabad 45320, Pakistan.
² Department of Biological Sciences, National University of Medical Sciences, Rawalpindi, Pakistan.
³ Department of Botany, Bacha Khan University, Charsadda, Khyber Pakhtunkhwa, Pakistan.
⁴ Department of Plant Sciences, Quaid-i-Azam University, Islamabad 45320, Pakistan.
⁵ Clinical Laboratory Sciences Department, Turabah University College, Taif University, Taif 21995, Saudi Arabia.
⁶ Department of Biotechnology, College of Science, Taif University, P.O. Box 11099, Taif 21944, Saudi Arabia.

Abstract

The current study aims to investigate the anticancer potential of Periploca hydaspidis extracts against HCCLM3 and MDA-MB 231 cell lines with invasive properties and to identify molecular targets underlying its action mechanism. Cytotoxic screening of plant extracts was done via MTT assay against liver and breast cancer cell lines and GC/MS of the best cytotoxic fraction was performed to identify its chemical composition. Flow cytometry detected apoptosis and cell-cycle changes after drug treatment. The specified cells were studied for migration and invasion potential along with performing western blot analysis of proteins involved in apoptosis, cell-cycle, metastasis, and MAPK (Mitogen-activated protein kinase) cell-signaling pathway. The results revealed the crude methanol (PHM) fraction of P. hydaspidis shown dose and time dependent cell-proliferative inhibition response. GC/MS analysis detected 54 compounds of which fatty acids (29.8%), benzenoids (15.7%), and esters (14.3%) constituted the bulk. The inhibitory effect against cancer cells was linked with cell-cycle arrest at G0/G1 phase, induction of apoptosis, reduced migration and invasion capabilities post treatment. PHM induced apoptosis via downregulation of anti-apoptotic (survivin, B-cell lymphoma Extra-large; BCL-XL, X-linked inhibitor of apoptosis protein; XIAP, Myelocytomatosis; C-myc), metastatic (Matrix metallopeptidases 9/2; MMP9/2), and cell-cycle regulatory (cyclin D1 and E) proteins, whereas upregulation of pro-apoptotic proteins (Bcl-2 homologous antagonist/killer; BAK, Bcl-2-Associate X protein; BAX, cleaved caspases; 3,7,8,9, and PARP) and activation of MAPK (Jun amino-terminal kinase; JNK and P38) pathway. P38 was needed for PHM-induced apoptosis, where the inhibition of P38 by pharmacological inhibitor (SB239063) diminished the apoptotic effects. Overall, our results conclude that PHM can inhibit cell-proliferation and induce apoptotic effects by activation of P38 MAPK cell-signaling pathway. This suggests the methanol fraction of P. hydaspidis (PHM) to have anticancer compounds, potentially useful for treating liver and breast cancer. In future, one-step advance studies of PHM regarding its role in metastatic inhibition, immune response modulation for reducing tumor, and inducing apoptosis in suitable animal models would be an interesting and promising research area.

Keywords: GC/MS; HCCLM3; JNK; MAPK; MDA–MB 231; P38; PARP; Periploca hydaspidis.