Effects of partially purified fractions of kaffir lime (Citrus hystrix DC.) leaf on cell cycle in K562 and Molt4 leukemic cell lines

ผลของสารสกัดแยกบริสุทธิ์บางส่วนจากใบมะกรูด (Citrus hystrix DC.) ต่อวัฏจักรของเซลล์ ในเซลล์มะเร็งเม็ดเลือดขาวเพาะเลี้ยงชนิด K562 และ Molt4

Name: Wipavee Utthawang

ThaSH: Leukemia

Classification :.DDC: 616.99419

ThaSH: Cancer cells

Abstract: Leukemia is a hematopoietic disorder of hematopoietic stem cells in bone marrow and results in uncontrolled leukemic blood cell proliferation. Nowadays, chemotherapy is an effective method and is widely used for leukemia treatment. To avoid severe side effects of chemotherapeutic drugs, plants having anticancer activity are an alternative choice for leukemia patients. In this study, kaffir lime (Citrus hystrix DC.) leaf is a choice of Thai traditional medicine plant. This study aims to investigate anti-proliferative effects of partially purified fractions of kaffir lime leaf and to examine their activity on cell cycle arrest as well as cell cycle regulatory proteins at related phase after treatments in K562 (human chronic myelogenous leukemia) and Molt4 (human acute lymphoblastic leukemia) leukemic cell lines. The partially purified fraction No. 9, 10, and 11 (F9, F10, and F11) from hexane fractional extract were examined due to their anti-leukemic activity observed in a previous study. Therefore, the partial purified fraction No. 9, 10, and 11 were further examined their effects on cell cycle progression in K562 and Molt4 leukemic cell models by flow cytometry. The effects on regulatory proteins were determined by Western blot analysis. The cytotoxic effects of three purified fraction No. 9, 10, and 11 were studied by MTT assay. The result showed that three purified fractions showed cytotoxic effects on K562 and Molt4 cell lines with IC50 values ranging from 32.0 to 80.0 µg/mL in K562 cells and 20.8 to 24.5 µg/mL in Molt4 cells. The purified fraction No. 9 was selected for further experiment due to the good h effective in both leukemic cells. IC20 values (non-cytotoxic doses) of purified fraction No. 9 were used for cell cycle arrest study by flow cytometer and cell cycle protein control by Western blot analysis. The purified fraction No. 9 with the concentrations of 10.0 and 2.5 µg/mL could increase K562 and Molt4 cell populations and arrest both cells at G2/M phases by flow cytometer at 48 h, respectively. It also induced p53 level in K562 and Molt4 cells. This phenomenon indicated that cell cycle progression was inhibited at G2/M phases. Moreover, it inhibited cyclin A, cyclin B, and cdc2 protein expression in a time- and dose-dependent manner in both cell lines. This study provides new knowledge of possible compound group in the partially purified hexane fractional extracts from kaffir lime leaf on cell cycle progression. The new knowledge from this study should be applied for drug development in complementary and alternative medicine for leukemia treatment in the future or co-treatment with chemotherapeutic drugs to reduce their side effects in leukemia patients.

Chiang Mai University Library

Address: Chiang Mai

Email: cmulibref@cmu.ac.th

Name: Chuda Chittasupho

Role: Cairman

Name: Songyot Anuchapreeda

Role: Member

Name: Siripogn Okqrogi

Role: Member

Name: Chadarat Ampasavate

Role: Member

Name: Sawitree Chiampanichayakul)

Role: Member

Created: 2016

Modified: 2020-01-06

Issued: 2020-01-06

วิทยานิพนธ์/Thesis

application/pdf

CallNumber: Th 616.99419 W797E

eng

DegreeName: Master of Science

Level: Master's Degree

Descipline: Medical Technology

Grantor: Chiang Mai University

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