Rapamycin synergizes with low dose oxaliplatin in the HCT116 colon cancer cell line by inducing enhanced apoptosis

Lu,Xueying; Wei,Haibo; Zhang,Xiaojin; Zheng,Wenxin; Chang,Cheng; Gu,Jinyu

doi:10.3892/ol.2011.299

Rapamycin synergizes with low dose oxaliplatin in the HCT116 colon cancer cell line by inducing enhanced apoptosis

Authors:
- Xueying Lu
- Haibo Wei
- Xiaojin Zhang
- Wenxin Zheng
- Cheng Chang
- Jinyu Gu
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Affiliations: Immunology Department, Basic Medical Science College, Harbin Medical University, Heilongjiang 150081, P.R. China, Department of General Surgery, The Second Affiliated Hospital of Harbin Medical University, Nangang, Harbin, Heilongjiang 150086, P.R. China
Published online on: May 9, 2011 https://doi.org/10.3892/ol.2011.299
Pages: 643-647

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Abstract

The present study aimed to examine the combined effects of oxaliplatin (L‑OHP) and rapamycin (RAPA) in the HCT116 colon cancer cell line. The growth inhibitory effect was evaluated by MTT assay as a monotherapy or combination therapy. IC50 values were determined using CalcuSyn 2.0 software. To determine the interaction of the drugs, the combination index (CI) was calculated using the Chou‑Talalay method. Apoptosis was investigated using flow cytometry and Western blotting. Acridine orange staining was employed to observe morphological changes. The results showed the IC50 values of L‑OHP and RAPA to be 8.35±0.78 µM (r=0.99) and 223.44±38.10 nM (r=0.94), respectively. CI was ≤1 when L‑OHP was used at doses ranging from 1 to 5 µM plus RAPA at a dose of 10 nM, suggesting synergistic or additive effects. CI was ≥1 when 100 nM RAPA was used in combination with low‑dose L‑OHP, showing additive to antagonistic effects. The combination of L‑OHP (1 µM) and RAPA (10 nM) induced 19.76% Annexin V‑positive cells, which was found to be higher than L‑OHP (11.45%, p<0.01) or RAPA (6.89%, p<0.01) alone. The cleaved PARP protein expression levels were highest after 48 h of combination treatment. Acridine orange staining showed typical bright red Acidic vesicular organelles in the RAPA group, whereas the green condensed chromatin in the apoptotic bodies was found in both the L‑OHP and combination groups. In conclusion, at a cytostatic concentration, RAPA was found to potentiate the anti‑tumor effects of low‑dose L‑OHP in the HCT116 colon cancer cell by inducing enhanced apoptosis.

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