Hexane partition from Annona crassiflora Mart. promotes cytotoxity and apoptosis on human cervical cancer cell lines
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Cervical cancer is the third most commonly diagnosed tumor type and the fourth cause of cancer-related death in females. Therapeutic options for cervical cancer patients remain very limited. Annona crassiflora Mart. is used in traditional medicine as antimicrobial and antineoplastic agent. However, little is known about its antitumoral properties. In this study the antineoplastic effect of crude extract and derived partitions from A. crassiflora Mart in cervical cancer cell lines was evaluated. The crude extract significantly alters cell viability of cervical cancer cell lines as well as proliferation and migration, and induces cell death in SiHa cells. Yet, the combination of the crude extract with cisplatin leads to antagonistic effect. Importantly, the hexane partition derived from the crude extract presented cytotoxic effect both in vitro and in vivo, and initiates cell responses, such as DNA damage (H2AX activity), apoptosis via intrinsic pathway (cleavage of caspase-9, caspase-3, poly (ADP-ribose) polymerase (PARP) and mitochondrial membrane depolarization) and decreased p21 expression by ubiquitin proteasome pathway. Concluding, this work shows that hexane partition triggers several biological responses such as DNA damage and apoptosis, by intrinsic pathways, and was also able to promote a direct decrease in tumor perimeter in vivo providing a basis for further investigation on its antineoplastic activity on cervical cancer.
KeywordsAnnona crassiflora Mart Natural compounds Hexane partition apoptosis Cytotoxity and cervical cancer cell lines
Viviane A O Silva designed all the experiments as well as participated of data acquisition, its interpretation and drafted the manuscript. Viviane A O Silva, Ana Laura V Alves and Marcela N Rosa carried out the studies of cell culture including cytotoxicity and proliferation assay, wound healing migration assay, colony formation assay, and statistical analysis from Hexane partition. Viviane A O Silva, Marcela N Rosa and Larissa R V Silva carried out the studies of cell culture including cytotoxicity and proliferation assay, wound healing migration assay, drug combination studies and statistical analysis from crude extract. Aline Tansini helped to carry out flow cytometry assays and its interpretation. Matias Melendez, Giovanna Longato and Olga Martinho contributed to design some experiments, interpretation of data and involved in revising critically the manuscript. Ana Laura V Alves and Fernanda Cury helped to design and performed the in vivo experiments. Bruno G. Oliveira, Fernanda E. Pinto and Wanderson Romãohas been responsible for the preparation of extracts. Izabela Faria Gomes contributed to design some experiments and analysed about the profile of secondary compounds identified in the hexane partition. Rosy Ribeiro conceived the extracts and partition, participated in its design, interpretation of data. Rui Manuel Reis conceived the study, participated in its design and coordination, interpretation of data, drafted the manuscript and have been involved in revising it critically for important intellectual content. All authors read and approved the final manuscript.
This study was supported by grants from the FINEP (MCTI/FINEP/MS/SCTIE/DECIT-01/2013 - FPXII-BIOPLAT), CAPES, FAPEMIG, UFSJ and Barretos Cancer Hospital, all from Brazil.
Compliance with ethical standards
Conflict of interest
The authors confirm that this article content has no conflicts of interest.
All applicable international, national, and/or institutional guidelines for the care and use of animals were followed. All procedures performed in studies involving animals were in accordance with the ethical standards of the institution or practice at which the studies were conducted. This article does not contain any studies with human participants performed by any of the authors.
Informed consent was obtained from all individual participants included in the study.
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