95P - Anticancer activity of Inula recemosa root extract in human liver cancer cell line by attenuation of OCT4/Sox2 axis

Background

Hepatocellular carcinoma (HCC) has become one of the leading causes of death globally in males and is also rising in females at an alarming rate. The growth, oncogenicity, metastasis and therapeutic resistance of hepatocellular carcinoma is maintained by hepatic cancer stem cells (HCSCs). The development of new therapeutic approaches specifically targeting hepatic cancer stem cells using herbal medicine could propose new hope for advanced HCC treatment. To achieve therapeutic success, emphasis should also depend on inhibitors that would not only be more effective on tumors but have minimal normal tissue toxicity. Considering the hepatoprotective role of Inula recemosa suggested before, the aim of our present study was to understand the role of Inula recemosa root (IRE) on hepatocellular carcinoma, and hepatic cancer stemness.

Methods

The cytotoxicity effect of n-hexane extract of Inula recemosa was evaluated by MTT assay on HepG2 liver cancer and WRL68 liver normal cell lines. Colony formation was also performed to measure reproductive integrity after treating with different concentration over a prolonged period of time. Flowcytometry method was used for cell cycle analysis, apoptosis and the production of reactive oxygen species (ROS) by DCFDA. Effect of IRE on cancer stemness transcription factors like SOX 2 and OCT4 transcript level was determined by RT-PCR method. Immunocytochemistry method was used to check the change in cancer stemness marker protein expression after IRE treatment.

Results

IRE showed very low toxicity on WRL68 normal liver cell line but robust antiproliferative effect on HepG2 cell line by induction of apoptosis. The induction of apoptosis was supported by increase in ROS production level in IRE treatment groups when compared with untreated cells. Cell cycle analysis showed arrest in Sub G0 phase. IRE also attenuated transcript level and protein expression of stemness markers.

Conclusions

Our data endorses the potential of IRE in hepatocellular carcinoma on targeting the cancer stem cell transcription factors for the first time. We can conclude that IRE might open up new therapeutic avenues on advanced therapies of hepatocellular carcinoma in near future.

Clinical trial identification

Editorial acknowledgement

Legal entity responsible for the study

The authors.

Funding

Has not received any funding.

Disclosure

All authors have declared no conflicts of interest.