Project's information

Project's title Study on the molecular effect of secondary compounds extracted from Helicteres hirsuta Lour. on liver cancer cells
Project’s code GUST.STS.ĐT-2020-SH03
Research hosting institution Graduate University of Science and Technology
Project leader’s name Dr. Nguyen Van Tru
Project duration 01/06/2020 - 30/06/2022
Project’s budget 300 million VND
Classify Fair
Goal and objectives of the project

- Evaluation of biological activity at the in vitro level of crude extracts with different extraction solvents and purified compounds on liver cancer cell lines.
- Studying the effect at the molecular level of EMT marker (mRNA, proteins) treated secondary compounds isolated from Helicteres hirsuta Lour. on liver cancer cell lines.

Main results

1. The secondary compounds have been successfully isolated from Helicteres hirsuta Lour. The total methanol extraction obtained accounted for 5.07% of the initial dry mass. Layered extraction with solvents of different polarity, recorded the highest percentage of Dichloromethane extract. Followed by n-Hexane and ethyl acetate extracts.
2. Analyzed 20 substances present in extracts from alfalfa plant by gas chromatography coupled mass spectrometry. Purification of 4 main compounds from the extract of Helicteres hirsuta Lour. from the solvent was dichloromethane/ The purified compound were confirmed by HPLC.
3. Diclomethane extract had the highest cytotoxic activity on liver cancer cell lines. IC50 growth inhibition value recorded IC50 = 150.11±1.78 µg/ml from diclomethano extract on Huh7 liver cancer cell line, on HepG2 cancer cell line IC50 = 113.11±2.77 µg/ml. In addition, the clean substance Hexadecanoic acid, the main ingredient in the diclomethano extract, showed the best cytotoxic activity on the studied cancer cell lines. The growth inhibition value of 50 % IC50 of Hexadecanoic acid on liver cancer cell lines Hepg2, Chang cell and Huh7 recorded IC50 as 112.01±1.58 µg/ml, 182.28 ±3.18, and 134.64±2.78 µg/ml respectively. 
4. Hexadecanoic acid and diclomethan extract showed significant inhibitory activity on wound healing assay on liver cancer cell lines such as Hepg2, Huh7 and Chang cell. In addition, the Hexadecanoic acid also showed higher inhibitory activity on wound healing than the diclomethane extract on studied liver cancer cell lines.
5. Hexadecanoic acid and diclomethane extract were also reported to inhibite  colony formation and colony area with liver cancer cell lines such as Hepg2, Huh7 and Chang cell at a treatment concentration of 50 µg/ml of Hexadecanoic acid with cell colony forming area ratios of 53.88, 59.78 and 65.60, respectively. Hexadecanoic acid also gave a higher inhibitory effect than the crude extract of diclomethane, at the same time.
6. The the inhibitory activity of migration and invasion assay on liver cancer cell lines was also observed when treated with hexadecanoic acid and dichloromethane extract. The number of invasived cells when treated with Hexadecanoic acid at a concentration of 50 µg/ml in the liver cancer cell lines Hepg2, Huh7 and Chang cell was recorded as 53.88, 67.23 and 70.42, respectively, when compared with the group. In addition, Hexadecanoic acid showed the inhibit invasive metastasis higher than that of the dichloromethane extract.
7. The expression of several genes that related to cell metastasis such as Snail, Slug, Twist, Zeb, and E-Cadherin was significantly reduced by the treatment of the Hexadecanoic acid and the extract of diclomethane, and showed dependence manner at both the mRNA and protein levels. In contrast, the purified Hexadecanoic acid and diclomethane extract enhanced N-Cadherin protein expression at both mRNA and protein levels.

Novelty and actuality and scientific meaningfulness of the results

- Develop experiments to help screen anticancer activity for new compounds.
- Evaluate the expression level of EMT markers related to the invasive metastasis of liver cancer cells when treated with clean substance and extract from Ansa plant.
- The project has published 2 pape.

Products of the project
- Scientific papers in referred journals (list): 02 Papers
Tru Van Nguyen   Luong Thi Tran  Huong Ho Thi  Ho Manh Tuong  Van Hong Nguyen  Van Hanh Nguyen  Thuy Thi Bich Le (2022). Ethyl Acetate Extract of Helicteres hirsuta Suppresses MCF-7 Human Breast Cancer Cell Mobility. Annual Research & Review in Biology, 10-17 DOI: 10.9734/arrb/2022/v37i730517 
Tru Van Nguyen, Luong Thi Tran, Huong Thi Ho, Thi Thao Do, Van Hanh Nguyen, Duc Hoang Le, Ngoc Anh Ho, Thi Nguyet Doan, Duong Hoang Nguyen, Duong Pham Quang, Thuy Thi Bich Le. Dichloromethane Extract of Helicteres hirsuta Inhibits Human Hepatocellular Carcinoma Cell Proliferation and Migration Via Downregulating EMT Marker Expressions. Latin American Journal of Pharmacy
- Patents (list): 01 utility solution
Research area

This research direction should continue to contribute effectively to the conservation of other endangered species in Vietnam
It is necessary to continue to investigate the hepatocellular anticancer activity of  Hexadecanoic acid extracted from Helicteres hirsuta L. in vivo level.

Images of project