The effect of commelina benghalensis (commelinaceae) on microrna expression in cervical cancer cells

Abstract

Cervical cancer is a health burden in many countries worldwide and is the most prevalent cancer in women across the African continent. Even with the current treatment strategies such as radiotherapy, chemotherapy and surgery, cervical cancer still causes high rates of morbidity and mortality. Novel treatment strategies for this cancer are urgently needed. Medicinal plants are gaining a lot of interest as sources of bioactive compounds for the treatment of different diseases, including cancer. Commelina benghalensis extracts have been shown to have anti-cancer properties against Jurkat T cells and alveolar epithelial cells (AEC). However, there has been no report on the activity of this plant against cervical cancer cells. Additionally, microRNA (miRs) have shown to regulate carcinogenesis during the development of cervical cancer cells, but the effects of C. benghalensis extracts on miRs have not previously been investigated. The influence of C. bengalensis acetone extract on cell metabolism, apoptosis and miRNA expression in HeLa cervical cells were tested in this study. The stem and leaf extracts of C. bengalensis decreased the metabolic activity in HeLa cells as shown by MTT assay and caused cell apoptosis as demonstrated with Annexin V flow cytometry. Twelve miRs were identified that are involved in the apoptosis pathway and selected for further studies. HeLa cells were treated with 400 and 500 μg/ml C. benghalensis leaf and stem extracts for 24 hours. Six miRs were significantly upregulated by the different extracts and concentrations, namely miR-7, miR-20a, miR-34a, miR-182, miR-143 and miR-200a. Six of the miRs were not significantly affected by the C. bengalensis extracts, specifically miR-21, miR-27b, miR-29a miR-99a, miR-99b and miR-497. None of the miRs were down regulated by the C. benghalensis extracts. The 400 μg/ml stem and leaf extracts, as well as the 500 μg/ml leaf extract upregulated some miRs that are usually already upregulated in cervical cancer cells, thereby potentiating carcinogenesis. Following this observation, it can be concluded that these extracts may not be good candidates for the treatment of cervical cancer. The 500 μg/ml stem extract was the best candidate for potential cancer treatment as it significantly upregulated only miR-34a which is usually downregulated in cervical cancer. This concentration can be further investigated for the treatment of cervical cancer.

Wêreldwyd het sevikale kanker ‘n hoë siektelas en is die algemeenste kanker onder vrouens in Afrika. Selfs met die huidige behandeling vir servikale kanker met chemoterapie, radioterapie en chirugie het diè kanker steeds ‘n hoë mortaliteit en morbiditeit insidensie. Nuwe behandelingsmodaliteite word dringend vir servikale kanker benodig. Plant ekstrakte van Commelina benghalensis het in Jurkat T-selle en EAC selle teen-kanker einskappe getoon. MikroRNAs het ‘n effeck op die karsinogenese van servikale kanker, maar die effek van C. benghalensis op mikroRNAs in servikale kanker is nog nie getoets nie. In die navorsingsprojek word die effek van C. benghalensis asetoon plant ekstrakte op sel metabolisme, apoptose en die microRNA uitdrukking in HeLa selle ondersoek. Daar is gevind dat die stam en blaar ekstrakte van C. benghalenis die metabolisme in HeLa selle verlaag met die gebruik van MTT-toets, asook om apoptose te induseeer wat bewys is met Annexin V vloeisitometrie. Twaalf mikroRNAs wat die apoptose pad beinvloed is geidentifiseer vir verdere studie. HeLa selle is vir 24 uur met 400 μg/ml en 500μg/ml stam en blaar esktrakte behandel, waarna die mikroRNA uitdrukking bepaal is. Ses mikroRNA was opgereguleer, naamlik: miR-7, miR-20a, miR-34a, miR-182, miR-143 en miR-200a. Die ander ses mikroRNA, naamlik miR-21, miR-27b, miR-29 miR-99a, miR-99b, en miR-497 was nie betekenisvol geafekkteer deur die C. benghalensis blaar en stam ekstrakte nie. Geen van die mikroRNAs was afgereguleer deur die C. benghalensis ekstrakte nie. Die 400 μg/ml stam en blaar ekstrakte sowel as die 500μg/ml blaar ekstrakt het microRNAs opgereguleer, wat reeds in servikale kanker ook opgereguleer is en kan dus verder karsinogenese ondersteun. In die lig hiervan is diè konsentrasies nie goeie kandidate vir servikale kanker behandeling nie. Die beste kandidaat vir potensiële servikale kanker behandeling was die 500μg/ml stam ekstrak wat slegs miR-34 opgereguleer wat gewoonlik verlaag is in servikale kanker.

Umdlavuza womlomo wesibeletho ungumthwalo wezempilo emazweni amaningi emhlabeni jikelele futhi uwumdlavuza odlange kakhulu kwabesifazane ezwenikazi lonke lase-Afrika. Ngisho namasu amanje okwelapha afana neradiotherapy, chemotherapy kanye nokuhlinzwa, umdlavuza womlomo wesibeletho usabangela izinga eliphezulu lokugula nokufa. Amasu amasha okwelapha lo mdlavuza adingeka ngokushesha. Izitshalo zokwelapha zithola isithakazelo esikhulu njengemithombo ye-bioactive compounds yokwelapha izifo ezihlukahlukene, kuhlanganise nomdlavuza. Ukukhishwa kwe-Commelina benghalensis kukhonjiswe ukuthi kunezinto zokulwa nomdlavuza ngokumelene namaseli e-Jurkat T namaseli e-alveolar epithelial (AEC). Kodwa-ke, awukho umbiko mayelana nokusebenza kwalesi sitshalo ngokumelene namaseli omdlavuza womlomo wesibeletho. Ukwengeza, i-MicroRNA (miRs) ibonise ukulawula i-carcinogenesis ngesikhathi sokuthuthukiswa kwamangqamuzana omdlavuza womlomo wesibeletho, kodwa imiphumela ye-C. benghalensis extracts kuma-miRs ayizange iphenywe ngaphambilini. Umthelela we-C. bengalensis acetone extract on cell metabolism, apoptosis kanye nenkulumo ye-miRNA kumaseli omlomo wesibeletho we-HeLa ahlolwe kulolu cwaningo. Isiqu nesiqephu seqabunga se-C. bengalensis sehlise umsebenzi we-metabolic kumaseli e-HeLa njengoba kuboniswe ukuhlolwa kwe-MTT futhi kwabangela i-apoptosis yeseli njengoba kuboniswe nge-Annexin V Flow Cytometry. Ama-miR ayishumi nambili ahlonziwe abandakanyeka kumzila we-apoptosis futhi akhethelwa ukuqhubeka nezifundo. Amaseli e-HeLa aphathwe nge-400 kanye ne-500 μg/ml C. Iqabunga le-Benghalensis ne-stem extract amahora angu-24. Ama-miR ayisithupha aye alawulwa kakhulu izingcaphuno ezihlukene nokugxiliswa kuzo, okuyi-miR-7, miR-20a, miR-34a, miR-182, miR-143 kanye ne-miR-200a. Ama-miR ayisithupha awazange athintwe kakhulu izingcaphuno ze-C. bengalensis, ikakhulukazi i-miR-21, miR-27b, miR-29a miR-99a, miR-99b ne-miR-497. Awekho ama-miR aye phansi alawulwa yi-C. benghalensis extracts. I-400 μg/ml isiqu kanye nezikhishiwe zeqabunga, kanye ne-500 μg/ml yeqabunga elikhishwe kulawula amanye ama-miR ngokuvamile asevele elawulwa kumaseli omdlavuza womlomo wesibeletho, ngaleyo ndlela enze i-carcinogenesis. Ukulandela lokhu kubheka, kungaphethwa ngokuthi lezi zingcaphuno zingase zingabi abantu abafanelekile ekwelapheni umdlavuza womlomo wesibeletho. Ukukhishwa kwesiqu okungu-500 μg/ml bekuyikhandidethi elingcono kakhulu lokwelapha umdlavuza okungenzeka ukuthi lilawule kakhulu i-miR-34a kuphela evamise ukwehliswa kumdlavuza womlomo wesibeletho. Lokhu kugxilwa kungaphenywa kabanzi ekwelapheni umdlavuza womlomo.