Published online Feb 7, 2014. doi: 10.3748/wjg.v20.i5.1348
Revised: November 6, 2013
Accepted: December 5, 2013
Published online: February 7, 2014
Processing time: 164 Days and 4.4 Hours
AIM: To investigate H2O2-induced promotion proliferation and malignant transformation in WB-F344 cells and anti-tumor effects of ursolic acid (UA) and oleanolic acid (OA).
METHODS: WB-F344 cells were continuously exposed to 7 x 10-7 mol/L H2O2 for 21 d. Observations of cell morphology, colony formation rates, flow cytometric analysis of cell cycle changes and aneuploidy formation indicated that H2O2 was able to induce malignant transformation of WB-F344 cells. We treated malignantly transformed WB-F344 cells with 4 μmol/L OA or 8 μmol/L UA for 72 h and analyzed the cell cycle distribution by flow cytometry.
RESULTS: MTT assay showed that 7 x 10-7 mol/L H2O2 decreased G1 phase subpopulation from 73.8% to 49.6% compared with the control group, and increased S phase subpopulation from 14.5% to 31.8% (P < 0.05 vs control group). Cell morphology showed that nucleus to cytoplasm ratio increased, many mitotic cells, prokaryotes and even tumor giant cells were shown in H2O2-induced WB-F344 cells. Fluorescence activated cell sorting analysis showed that WB-F344 cell aneuploidy increased to 12% following H2O2 treatment. Flow cytometric analysis of the transformed WB-F344 cells following treatment with OA (4 μmol/L) and UA (8 μmol/L) showed that OA increased G1 subpopulation to 68.6%, compared to 49.7% in unexposed cells. UA increased G1 subpopulation to 67.4% compared to 49.7% in unexposed cells (P < 0.05 vs H2O2 model group).
CONCLUSION: H2O2 causes the malignant transformation of WB-F344 cells. OA and UA exert anti-tumor effects by inhibiting the proliferation in malignantly transformed WB-F344 cells.
Core tip: It is known that H2O2 can promote tumorigenesis. Here, we used H2O2 as a premalignant and malignant agent to induce proliferation and malignant transformation in quiescent rat liver oval cell line WB-F344. Multistage carcinogenic processes provide the basis for interrupting and reversing precancerous changes; therefore, reversing precancerous changes is critical for tumor prevention and treatment. The salient and novel findings of the study are that ursolic acid (UA) and oleanolic acid (OA) induced malignantly transformed WB-F344 cell arrest in the G1 phase and apparently inhibited the proliferation of these cells. These results better our understanding of the antitumor effects of OA and UA.