SUBSCRIBE
Search JIM Advanced Search

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

RichHTML

2

PDF (PC)

82

Like

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
[1] Cai-lian Fana, Wan-jun Caib, Meng-nan Ye, Miao Chen, Yi Dai. Qili Qiangxin, a compound herbal medicine formula, alleviates hypoxia-reoxygenation-induced apoptotic and autophagic cell death via suppression of ROS/AMPK/mTOR pathway in vitro. Journal of Integrative Medicine, 2022, 20(4): 365-375.
[2] Hong-xiao Li, Ling Shi, Shang-jie Liang Chen-chen Fang, Qian-qian Xu, Ge Lu, Qian Wang, Jie Cheng, Jie Shen, Mei-hong Shen. Moxibustion alleviates decreased ovarian reserve in rats by restoring the PI3K/AKT signaling pathway. Journal of Integrative Medicine, 2022, 20(2): 163-172.
[3] Mohamed S. Othman, Sofian T. Obeidat, Amal H. Al-Bagawi, Mohamed A. Fareid, Alaa Fehaid, Ahmed E. Abdel Moneim. Green-synthetized selenium nanoparticles using berberine as a promising anticancer agent#br#
#br#
. Journal of Integrative Medicine, 2022, 20(1): 65-72.
[4] Hyongjun Jeon, Hee-Young Kim, Chang-Hwan Bae, Yukyung Lee, Sungtae Koo, Seungtae Kim. Korean red ginseng decreases 1-methyl-4-phenylpyridinium-induced mitophagy in SH-SY5Y cells. Journal of Integrative Medicine, 2021, 19(6): 537-544.
[5] Victor Abou Nehmi Filho, Gilson Masahiro Murata, Ruan Carlos Macêdo de Moraes, Gabriely Cristina Alves Lima, Danielle Araujo de Miranda, Katrin Radloff, Raquel Galvão Figuerêdo Costa, Joyce de Cassia Rosa de Jesus, Jéssica Alves De Freitas, Nayara Izabel Viana, Ruan Pimenta, Katia Ramos Moreira Leite, José Pinhata Otoch, Ana Flávia Marçal Pessoa. A novel supplement with yeast β-glucan, prebiotic, minerals and Silybum marianum synergistically modulates metabolic and inflammatory pathways and improves steatosis in obese mice. Journal of Integrative Medicine, 2021, 19(5): 439-450.
[6] Hao Gou, Ruo-chen Huang, Yong-hua Su, Wei Li. Design of dual targeting immunomicelles loaded with bufalin and study of their anti-tumor effect on liver cancer. Journal of Integrative Medicine, 2021, 19(5): 408-417.
[7] Porwornwisit Tritripmongkol, Tullayakorn Plengsuriyakarn, Mayuri Tarasuk, Kesara Na-Bangchang. In vitro cytotoxic and toxicological activities of ethanolic extract of Kaempferia galanga Linn. and its active component, ethyl-p-methoxycinnamate, against cholangiocarcinoma. Journal of Integrative Medicine, 2020, 18(4): 326-333.
[8] Varuni Colamba Pathiranage, Ira Thabrew, Sameera R. Samarakoon, Kamani H. Tennekoon, Umapriyatharshini Rajagopalan, Meran K. Ediriweera. Evaluation of anticancer effects of a pharmaceutically viable extract of a traditional polyherbal mixture against non-small-cell lung cancer cells. Journal of Integrative Medicine, 2020, 18(3): 242-252.
[9] Maged Mohamed Maher Abou-Hashem, Dina Mohamed Abo-elmatty, Noha Mostafa Mesbah, Ahmed Mohamed Abd EL-Mawgoud. Induction of sub-G0 arrest and apoptosis by seed extract of Moringa peregrina (Forssk.) Fiori in cervical and prostate cancer cell lines. Journal of Integrative Medicine, 2019, 17(6): 410-422.
[10] Yi Ran Hu, San Li Xing, Chuan Chen, Ding Zhu Shen, Jiu Lin Chen. Tiaoxin Recipe, a Chinese herbal formula, inhibits microRNA-34a expression in the APPswe/PS1ΔE9 mouse model of Alzheimer's disease. Journal of Integrative Medicine, 2019, 17(6): 404-409.
[11] Supakanya Kumkarnjana, Rutt Suttisri, Ubonthip Nimmannit, Apirada Sucontphunt, Mattaka Khongkow, Thongchai Koobkokkruad, Nontima Vardhanabhuti. Flavonoids kaempferide and 4,2′-dihydroxy-4′,5′,6′-trimethoxychalcone inhibit mitotic clonal expansion and induce apoptosis during the early phase of adipogenesis in 3T3-L1 cells. Journal of Integrative Medicine, 2019, 17(4): 288-295.
[12] Shriniwas S. Basaiyye, Sanjay Kashyap, Kannan Krishnamurthi, Saravanadevi Sivanesan. Induction of apoptosis in leukemic cells by the alkaloid extract of garden cress (Lepidium sativum L.). Journal of Integrative Medicine, 2019, 17(3): 221-228.
[13] Carlos R. Oliveira, Daniel G. Spindola, Daniel M. Garcia, Adolfo Erustes, Alexandre Bechara, Caroline Palmeira-dos-Santos, Soraya S. Smaili, Gustavo J.S. Pereira, André Hinsberger, Ezequiel P. Viriato, Maria Cristina Marcucci, Alexandra C.H.F. Sawaya, Samantha L. Tomaz, Elaine G. Rodrigues, Claudia Bincoletto. Medicinal properties of Angelica archangelica root extract: Cytotoxicity in breast cancer cells and its protective effects against in vivo tumor development. Journal of Integrative Medicine, 2019, 17(2): 132-140.
[14] Bruno José Martins Da Silva, Sandro Wilson Gomes Pereira, Ana Paula Drummond Rodrigues, José Luiz Martins Do Nascimento, Edilene Oliveira Silva. In vitro antileishmanial effects of Physalis angulata root extract on Leishmania infantum. Journal of Integrative Medicine, 2018, 16(6): 404-410.
[15] Eric Wei Chiang Chan, Siu Kuin Wong, Hung Tuck Chan . Casticin from Vitex species: A short review on its anticancer and anti-inflammatory properties. Journal of Integrative Medicine, 2018, 16(3): 147-152.
Viewed
Full text


Abstract

Cited
  Shared   
  Discussed   
[1] Wei-xiong Liang. Problems-solving strategies in clinical treatment guideline for traditional Chinese medicine and integrative medicine. Journal of Chinese Integrative Medicine, 2008, 6(1): 1-4
[2] Zhao-guo Li. Discussion on English translation of commonly used sentences in traditional Chinese medicine: part one. Journal of Chinese Integrative Medicine, 2008, 6(1): 107-110
[3] Jun Hu, Jian-ping Liu. Non-invasive physical treatments for chronic/recurrent headache. Journal of Chinese Integrative Medicine, 2008, 6(1): 31
[4] Xue-mei Liu, Qi-fu Huang, Yun-ling Zhang, Jin-li Lou, Hong-sheng Liu, Hong Zheng. Effects of Tribulus terrestris L. saponion on apoptosis of cortical neurons induced by hypoxia-reoxygenation in rats. Journal of Chinese Integrative Medicine, 2008, 6(1): 45-50
[5] . Uniform requirements for manuscripts submitted to biomedical journals: Writing and editing for biomedical publication (Chinese version, part two). Journal of Chinese Integrative Medicine, 2010, 8(11): 1001-1005
[6] Daniel Weber, Janelle M Wheat, Geoffrey M Currie. Inflammation and cancer: Tumor initiation, progression and metastasis,and Chinese botanical medicines. Journal of Chinese Integrative Medicine, 2010, 8(11): 1006-1013
[7] Hong Liu , Guo-liang Zhang, Li Shen , Zhen Zeng, Bao-luo Zhou, Cheng-hai Liu, Guang Nie . Application and evaluation of a pseudotyped virus assay for screening herbs for anti-H5Nl avian influenza virus. Journal of Chinese Integrative Medicine, 2010, 8(11): 1036-1040
[8] Zhao-guo Li . A discussion of English translation of 1995 and 1997 Chinese National Standards of Traditional Chinese Medical Terminologies for Clinical Diagnosis and Treatment. Journal of Chinese Integrative Medicine, 2010, 8(11): 1090-1096
[9] Rui Jin, Bing Zhang. A complexity analysis of Chinese herbal property theory: the multiple formations of herbal property (Part 1). Journal of Chinese Integrative Medicine, 2012, 10(11): 1198-1205
[10] Hui-min Liu, Xian-bo Wang, Yu-juan Chang, Li-li Gu. Systematic review and meta-analysis of randomized controlled trials of integrative medicine therapy for treatment of chronic severe hepatitis. Journal of Chinese Integrative Medicine, 2012, 10(11): 1211-1228
Journal of Integrative Medicine, ISSN: 2095-4964 CN 31-2083/R
Copyright © Journal of Integrative Medicine
Powered by Beijing Magtech Co. Ltd
沪公网安备31010602003416号      沪ICP备18017633号-3