(P13-023-23) Apoptotic Activity of Crude Extract From Spike of Stomopneustes variolaris on Cervical Adenocarcinoma HeLa Cell Line In Vitro

Objectives: To observe the anticancer potential of crude extract from spike of Stomopneustes variolaris in human cervical adenocarcinoma HeLa cell in vitro.

Methods: Cell viability in vitro used to show the IC₅₀ of the crude extract in treated HeLa cells. The signal proteins affecting the change in cell viability following the treatment were identified using Westernblot. The expression level of apoptotic genes that are essential for cell death was quantified using RT-qPCR. One of the features of the induction of apoptosis is the disintegration of nuclear DNA and the subsequent nucleosomal fragments. Hence, this study employed DNA fragmentation test using gel electrophoresis. GC-MS was used to identify some of potential components in the crude extract.

Results: The extract had shown cytotoxic effect in HeLa cells with an IC₅₀ of 723.1 ± 9.73 µg/ml, but no toxicity in HEK293 and HUVEC. Westernblot was used to detect protein expression; Bax, caspase-8, and IκBα were increased, whereas IKKα and p-NFκB-65(Ser 536) were downregulated. RNA/mRNA expression was revealed by RT-qPCR, and fold changes of caspase-3, cytochrome-c, Bax, Apaf-1, caspase-9, and Bak genes’ expression were raised. Using gel electrophoresis, the treatment groups showed more DNA fragmentation than the control group. Anticancer activity could be attributed to dodecanoic acid, hexadecanoic acid, and tetradecanoic acid content.

Conclusions: The result showed the potential use of the crude extract of a spike from S. variolaris with anticancer activity against cervical cancer.

Funding Sources: Pukyong National University