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Journal of Chinese Integrative Medicine ›› 2008, Vol. 6 ›› Issue (12): 1267-1274.doi: 10.3736/jcim20081212

• Original Experimental Research • Previous Articles     Next Articles

Antileukemic mechanism of resveratrol in vitro and in mice bearing L1210-tumor

Tan Li1(), Wei Wang2, Tong Li3   

  1. 1. Department of Immunology, Medical College of the Chinese People's Armed Police Force, Tianjin 300162, China
    2. Department of Nephrology, Affiliated Hospital, Medical College of the Chinese People's Armed Police Force, Tianjin 300162, China
    3. Department of Ultrasonography, Xinjiang Armed Police General Hospital, Urumqi, Xinjiang Uygur Autonomous Region 830000, China
  • Received:2008-07-10 Online:2008-12-20 Published:2008-12-15

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Objective: 

To elucidate the molecular mechanism of resveratrol against leukemia both in vitro and in vivo.


Methods: 

Three kinds of leukemia cell lines, HuT-78, Jurkat and L1210 cells, were used in this study. After different doses of Res treatment, methyl thiazolyl tetrazolium (MTT) colorimetry was used to detect the cell proliferation; apoptosis was detected by flow cytometry; Western blot and immuno-precipitation method were used to detect the Bcl-2 and Bax proteins expression and the activity of phospho-signal transducer and activator of transcription 3 (p-STAT3). Besides, a total of 40 BALB/c mice were randomly divided into untreated group and 12.5, 25 and 50 mg/(kg·d) Res groups. Then, leukemia-bearing model was established by L1210 cells subcutaneous injection. Interleukin-6 (IL-6) was detected to determine the secretion function of T lymphocytes of the mice; the IL-6 mRNA expression in the liver tissue of mice was also detected by reverse transcription-polymerase chain reaction (RT-PCR), and the expression of the p-STAT3 protein was measured by Western blot and immunohistochemical method.


Results: 

The results indicated that resveratrol could inhibit the proliferation of HuT-78, Jurkat and L1210 cells and significantly induce the cell apoptosis. At the same time, the radio of Bcl-2/Bax and expression of p-STAT3 protein were decreased either. Furthermore, resveratrol could reduce the expression of IL-6 mRNA and intracellular content of IL-6, and decrease the expression of p-STAT3 protein in the liver of leukemic mice at a dose-dependent manner.


Conclusion: 

Resveratrol can function as an antileukemic agent through inducing apoptosis, modulating IL-6 and STAT3 both in vitro and in vivo.

Key words: resveratrol, apoptosis, leukemia, interleukin-6, STAT3 transcription factor, signal transducing adaptor proteins, mice, in vitro study

Table 1

Growth inhibition of resveratrol to leukemia cells HuT-78, Jurkat and L1210 in vitro (x±s, %)"

Group n Growth inhibition rate
12 h 24 h 48 h
Jurkat
6.25 μmol/L Res 3 9.91±0.40 15.13±1.60△ 22.77±0.91△△
12.5 μmol/L Res 3 15.07±1.82** 20.57±0.91**△△ 30.14±1.47**△△
25 μmol/L Res 3 20.18±2.12** 25.67±1.52**△△ 38.71±2.11**△△
50 μmol/L Res 3 25.25±1.35** 31.26±0.77**△△ 42.44±1.76**△△
100 μmol/L Res 3 30.18±2.26** 38.54±0.95**△ 45.31±1.53**△△
200 μmol/L Res 3 38.25±1.08** 58.16±1.35**△△ 63.58±1.33**△△
L1210
6.25 μmol/L Res 3 10.18±0.26 11.63±1.18 30.71±0.10△△
12.5 μmol/L Res 3 14.72±1.74** 18.74±1.22**△△ 35.42±2.07**△△
25 μmol/L Res 3 15.79±2.24** 32.47±1.79**△△ 43.34±1.60**△△
50 μmol/L Res 3 22.64±2.17** 40.50±2.17**△△ 48.80±1.21**△△
100 μmol/L Res 3 28.89±1.20** 52.35±1.13**△△ 62.17±1.98**△△
200 μmol/L Res 3 35.58±2.04** 61.04±1.95**△△ 70.53±0.80**△△
HuT-78
6.25 μmol/L Res 3 10.24±0.92 11.26±1.31 14.44±1.03△△
12.5 μmol/L Res 3 18.08±1.13** 20.10±1.46** 25.98±2.45**△
25 μmol/L Res 3 21.77±1.58** 33.35±1.83**△△ 48.40±1.72**△△
50 μmol/L Res 3 25.33±2.03** 40.96±1.27**△ 60.62±1.54**△△
100 μmol/L Res 3 35.03±1.59** 47.89±1.95**△△ 66.60±1.19**△△
200 μmol/L Res 3 40.12±1.95** 61.38±1.65**△△ 82.14±1.67**△△

Figure 1

Effects of 100 μmol/L Res on apoptosis of leukemia cells after 24-hour treatment in vitro detected by flow cytometry The apoptotic and necrotic cell distribution is analyzed by Annexin V binding and PI uptake. Positioning of quadrants on Annexin V/PI dot plots is performed, and living cells (Annexin V-/PI-), early apoptotic/primary apoptotic cells (Annexin V+/PI-), late apoptotic/secondary apoptotic cells (Annexin V+/PI+) and necrotic cells (Annexin V-/PI+) are distinguished. Therefore, the total apoptotic proportion includes the percentage of cells with fluorescence Annexin V+/PI- and Annexin V+/PI+. A, C, E: HuT-78, Jurkat and L1210 cells treated with 100 μmol/L resveratrol; B, D, F: Control of HuT-78, Jurkat and L1210 cells."

Figure 2

Effects of 100 μmol/L Res on apoptosis of leukemia cells after 12-, 24-, 48-hour treatment in vitro detected by flow cytometry **P<0.01, vs Jurkat cells; △△P<0.01, vs L1210 cells. n=3."

Figure 3

Effects of Res on expressions of Bcl-2 and Bax proteins in leukemia cells after 24-hour treatment in vitro detected by Western blotting 1: HuT-78 control group; 2: Jurkat control group; 3: L1210 control group; 4: HuT-78 IC50 Res group; 5: Jurkat IC50 Res group; 6: L1210 IC50 Res group."

Figure 4

Effects of Res on expressions of p-STAT3 and STAT3 proteins in leukemia cells after 24-hour treatment in vitro detected by immuno-precipitation method 1: Blank control group; 2: 25 μmol/L Res group; 3: 50 μmol/L Res group; 4: 100 μmol/L Res group."

Figure 5

Effects of Res on the expression of IL-6 mRNA in T lymphocytes in mice detected by RT-PCR 1: Blank control group; 2: 12.5 mg/(kg·d) Res group; 3: 25 mg/(kg·d) Res group; 4: 50 mg/(kg·d) Res group."

Figure 6

Content of IL-6 in T lymphocytes in mice detected by flow cytometry Positioning of quadrants on FITC/PE dot plots was performed and intracellular IL-6 (FITC+/PE+) were determined. A: Blank control group; B: 12.5 mg/(kg·d) Res group; C: 25 mg/(kg·d) Res group; D: 50 mg/(kg·d) Res group. *P<0.05, **P<0.01, vs blank control group. n=3."

Figure 7

Effects of Res on expression of p-STAT3 protein in liver tissue in mice detected by immunohistochemistry (Light microscopy, ×400) A: Blank control group; B: 12.5 mg/(kg·d) Res group; C: 25 mg/(kg·d) Res group; D: 50 mg/(kg·d) Res group."

Figure 8

Effects of Res on expression of p-STAT3 protein in liver tissue in mice detected by Western blotting 1: Blank control group; 2: 12.5 mg/(kg·d) Res group; 3: 25 mg/(kg·d) Res group; 4: 50 mg/(kg·d) Res group."

[1] Jang M, Cai L, Udeani GO , et al. Cancer chemopreventive activity of resveratrol, a natural product derived from grapes[J]. Science, 1997,275(5297):218-220
doi: 10.1126/science.275.5297.218 pmid: 8985016
[2] Yu HB, Pan CE, Wu WJ , et al. Effects of resveratrol on matrix metalloproteinase-9 expression in hepatoma cells[J]. Zhong Xi Yi Jie He Xue Bao, 2008,6(3):270-273
余海波, 潘承恩, 吴武军 , 等. 白藜芦醇对肝癌细胞基质金属蛋白酶-9表达的影响[J]. 中西医结合学报, 2008,6(3):270-273
[3] Fulda S, Debatin KM . Resveratrol modulation of signal transduction in apoptosis and cell survival: a mini-review[J]. Cancer Detect Prev, 2006,30(3):217-223
doi: 10.1016/j.cdp.2006.03.007
[4] Zhou YP . Randomized block design with Excel software in drug toxicology research[J]. Yao Xue Jin Zhan, 2005,29(9):425-427
周一平 . 用Excel软件进行药物毒理实验的随机分组[J]. 药学进展, 2005,29(9):425-427
[5] Wang Q, Li H, Liu N , et al. Correlative analyses of notch signaling with resveratrol-induced differentiation and apoptosis of human medulloblastoma cells[J]. Neurosci Lett, 2008,438(2):168-173
doi: 10.1016/j.neulet.2008.04.012
[6] Radogna F, Cristofanon S, Paternoster L , et al. Melatonin antagonizes the intrinsic pathway of apoptosis via mitochondrial targeting of Bcl-2[J]. J Pineal Res, 2008,44(3):316-325
doi: 10.1111/j.1600-079X.2007.00532.x
[7] Li Y, Suo XH, Niu ZY . The changes and clinical significance of levels of IL-2 and IL-6 in pre and post chemotherapy of acute leukemia patients[J]. Zhongguo Shi Yan Zhen Duan Xue, 2007,11(3):324-326
doi: 10.3969/j.issn.1007-4287.2007.03.016
李焱, 索晓慧, 牛志云 . 急性白血病患者化疗前后IL-2及IL-6水平的变化及临床意义[J]. 中国实验诊断学, 2007,11(3):324-326
doi: 10.3969/j.issn.1007-4287.2007.03.016
[8] Yin HQ, Qiao ZH, Zhu L , et al. Levels of intracellular IL-6 and IFN-γ in children with acute lymphoblastic leukemia[J]. Zhongguo Dang Dai Er Ke Za Zhi, 2006,8(6):461-463
doi: 10.3969/j.issn.1008-8830.2006.06.006
阴怀清, 乔振华, 朱镭 , 等. 儿童急性淋巴细胞性白血病细胞内IL-6、IFN-γ水平变化及其临床意义[J]. 中国当代儿科杂志, 2006,8(6):461-463
doi: 10.3969/j.issn.1008-8830.2006.06.006
[9] Takada Y, Bhardwaj A, Potdar P , et al. Nonsteroidal anti-inflammatory agents differ in their ability to suppress NF-kappaB activation, inhibition of expression of cyclooxygenase-2 and cyclin D1, and abrogation of tumor cell proliferation[J]. Oncogene, 2004,23(57):9247-9258
doi: 10.1038/sj.onc.1208169
[10] Yu LJ, Wu ML, Li H , et al. Inhibition of STAT3 expression and signaling in resveratrol-differentiated medulloblastoma cells[J]. Neoplasia, 2008,10(7):736-744
doi: 10.1593/neo.08304 pmid: 18592012
[11] Wung BS, Hsu MC, Wu CC , et al. Resveratrol suppresses IL-6-induced ICAM-1 gene expression in endothelial cells: effects on the inhibition of STAT3 phosphorylation[J]. Life Sci, 2005,78(4):389-397
doi: 10.1016/j.lfs.2005.04.052
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