Abstract
Globally, lung cancer accounts for 18% of cancer-associated mortalities. Among the subtypes, non-small cell lung cancer (NSCLC) is the most prevalent. The increased resistance and poor survival rates signify disease aggressiveness and thus require a search for an alternative anticancer molecule. Earlier, the sesquiterpene, i.e., compound 3 ((E)-methyl 6-acetoxy-7-methoxy-1-(2-methylpropylidene)-1H-indene-3-carboxylate) from Polygonum barbatum, was isolated, characterized by us, and reported for preliminary anticancer activity. Therefore, based on these results, this study was designed to explore the underlying molecular mechanism of apoptosis and metastasis against NCI-H460 cells. The molecular mechanism of compound 3 inducing apoptosis and inhibiting metastasis was elucidated by analyzing mitochondrial membrane potential, DNA fragmentation, clonogenic assay, invasion assay, and expression of apoptotic (caspases 3, 6, 8, 9, and BAK) and metastatic markers (MMP 2, MMP 9, and osteopontin). Compound 3 significantly inhibited cell proliferation and induced apoptosis via the intrinsic route, i.e., the mitochondrial pathway, by disrupting mitochondrial membrane potential. The enhanced expression of caspases 6, 9, BAK, and HRK with downregulation of Bcl-2L1 and Ki67 further confirmed the involvement of the intrinsic apoptotic pathway. Moreover, compound 3 restricted the invasive nature of NCI-H460 cells evinced by reduced cell invasion in Boyden chamber invasion assay and downregulating the expression of metastatic markers, i.e., matrix metalloproteinase 2/9 and VEGF. It was also found to block osteopontin by negatively regulating its expression, a marker protein in cancer management. Conclusively, this sesquiterpene exhibited potent anticancer and antimetastatic activity and can be explored further as possible pharmacophores.
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Abbreviations
- CCCP:
-
Carbonyl cyanide m-chlorophenyl hydrazone
- CAM:
-
Chorioallantoic membrane
- DAPI:
-
4′,6-Diamidino-2-phenylindole
- EDD4:
-
Embryo development day 4
- MMP:
-
Matrix metalloproteinases
- NSCLC:
-
Non-small cell lung cancer
- OPN:
-
Osteopontin
- PBS:
-
Phosphate buffer saline
- VEGF:
-
Vascular endothelial growth factors
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The authors are grateful to International Center for Chemical and Biological Sciences for providing partial financial support for this work.
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BZ, AA, AS, RU, SR, and TAK: conceptualization, methodology, perform experiment, collect data, and first draft preparation; AK, UF: provided compound 3; AA and SAA: supervision, manuscript editing, and finalization; all authors read and approved the manuscript and all data were generated in-house and that no paper mill was used.
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Zehra, B., Ahmed, A., Khan, A. et al. Sesquiterpene from Polygonum barbatum disrupts mitochondrial membrane potential to induce apoptosis and inhibits metastasis by downregulating matrix metalloproteinase and osteopontin in NCI-H460 cells. Naunyn-Schmiedeberg's Arch Pharmacol 395, 987–1001 (2022). https://doi.org/10.1007/s00210-022-02256-w
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DOI: https://doi.org/10.1007/s00210-022-02256-w