Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2022 Feb 1;23(2):583-590.
doi: 10.31557/APJCP.2022.23.2.583.

Effect of Betulinic acid Extraction from Guava (Psidium guajava Linn.) Leaves Against Human Cholangiocarcinoma Cells

Rassameepen Phonarknguen  1 , Saksit Nobsathian  2 , Kanjana Assawasuparerk  3
Affiliations

Effect of Betulinic acid Extraction from Guava (Psidium guajava Linn.) Leaves Against Human Cholangiocarcinoma Cells

Rassameepen Phonarknguen et al. Asian Pac J Cancer Prev. .

Abstract

Background: Betulinic acid (BA), a pentacyclic triterpene glycoside extract from guava (Psidium guajava Linn.) leaves, displays a variety of biological activities which exhibit cancer therapeutic properties associated with cancer growth inhibition in various kinds of human cancer cells including brain, breast, colorectal, cervical, lung and prostate gland. However, the effects on human cholangiocarcinoma cells have not previously been reported. Current study, we evaluated the activity of BA against human cholangiocarcinoma (HuCCA) cells.

Methods: The cytotoxicity analysis was measured by using MTT assay on HuCCA and BHK-21 cells. Analysis of apoptosis was evaluated by using staining with Hoechst 33342 and quantitative real-time PCR.

Results: The BA (50-800 µg/mL) significantly reduced the viability of HuCCA cells in a dose-dependent action with 50% inhibitory concentration (IC50) of 92.45 µg/mL at 24 h. It also induced apoptosis signaling pathway, such as nuclear chromatin condensation and fragmentation. Quantitative real-time PCR analysis demonstrated that BA increased p53, Bax and caspase-3 expression whilst it decreased Bcl-2 expression in the HuCCA cells in a dose dependent manner.

Conclusion: BA can inhibit the HuCCA cell viability and induce apoptosis of neoplastic cells. This study indicates that BA has effective treatment for cholangiocarcinoma in vitro. Consequently, BA may be used as a novel therapeutic agent for the treatment of cholangiocarcinoma in the future.

Keywords: Apoptosis; betulinic acid; cholangiocarcinoma cell; guava leaves (Psidium guajava Linn.).

PubMed Disclaimer

Figures

Figure 1
Figure 1
NMR Spectrum of Betulinic Acid Extracted from Guava (Psidium guajava Linn.) Leaves. (A) The H1 NMR (500 MHz), (B) The 13C NMR spectrum (125 MHz) and (C) The DEPT-135 and DEPT-90
Figure 2
Figure 2
The Effect of BA on the Cell Viability by MTT Assay Examined at 24 h. (A) Cell viability of HuCCA and BHK-21 cell lines after treatment with BA. (B) Cell viability of HuCCA cells after treatment with BA compared with 5-FU. All values are presented in mean ± SD, *** p < 0.001
Figure 3
Figure 3
Apoptosis Involved Nuclear Condensation and Fragmentation on HuCCA Cells. After 24 h treatment with BA was stained with Hoechst 33342 and measured by fluorescence microscopy at 40x magnification. (A) Control and (B) BA treatment group. White arrows show condensed and fragmented nuclei as the characteristic of apoptosis
Figure 4
Figure 4
Gene Expression in HuCCA Cells after BA Treatment for 24 h. Gene expression levels of (A) p53, (B) Bax, (C) Caspase-3, and (D) Bcl-2 were examined and compared with untreated group. *** p < 0.001
Figure 5
Figure 5
Gene Expression in HuCCA Cells after BA Treatment for 24 h. The ratio of Bax and Bcl-2, was examined and compared with untreated group. *** p < 0.001
See this image and copyright information in PMC

References

    1. Alakurtti S, Mäkelä T, Koskimies S, Yli-Kauthaluoma J. Pharmacological properties of the ubiquitous natural product botulin. Eur J Pharm Sci. 2006;29:1–13. - PubMed
    1. Anand V, Manikandan , Kumar V, et al. Phytopharmacological overview of Psidium gujava Linn. Phcog J. 2016;8:314–20.
    1. Baek MY, Cho JG, Lee DY, et al. Isolation of Triterpenoids from the Stem Bark of Albizia julibrissin and Their Inhibition Activity on ACAT-1 and ACAT-2. J Korean Soc Appl Biol Chem. 2010;53:310–15.
    1. Battle TE, Arbiser J, Frank DA. The natural product honokiol induces caspase-dependent apoptosis in B-cell chronic lymphocytic leukemia (B-CLL) cells. Blood. 2005;106:690–97. - PubMed
    1. Bras M, Queenan B, Susin SA. Programmed cell death via mitochondria: different modes of dying. Biochemistry (Mosc) 2005;70:231–9. - PubMed

LinkOut - more resources