Search JIM Advanced Search

Journal of Chinese Integrative Medicine ›› 2011, Vol. 9 ›› Issue (12): 1353-1359.doi: 10.3736/jcim20111212

• Original Experimental Research • Previous Articles     Next Articles

Medicated serum prepared with Chinese herbal medicine Zhizhen Recipe down-regulates activity of nuclear factor-κB and expression of P-glycoprotein in human colorectal cancer multidrug-resistant cell line HCT-8/VCR

Song-bai Cai, Xu Zhang, Zhi-xia Chen, Ling-chun Kong, Song-po Wang()   

  1. Department of Traditional Chinese Medicine, Shanghai First People's Hospital, Shanghai Jiao Tong University, Shanghai 200080, China
  • Received:2011-08-15 Accepted:2011-09-05 Online:2011-12-20 Published:2011-12-15

Objective: To investigate the effects of medicated serum prepared with Chinese herbal medicine Zhizhen Recipe (ZZR) on activity of nuclear factor-κB (NF-κB) and expression and function of P-glycoprotein (P-gp) in human colorectal cancer multidrug-resistant cell line HCT-8/VCR.
Methods: The multidrug resistance of HCT-8/VCR cells was detected by cell counting kit-8 method, and the experimental concentrations of ZZR-medicated serum were determined by the same way. HCT-8 and HCT-8/VCR cells were treated with ZZR-medicated serum of medium dose for 24 h. The activity of NF-κB was determined by enzyme-linked immunosorbent assay. The intracellular distribution of P-gp was detected by laser scanning confocal microscopy, and the mean fluorescence intensity of rhodamine 123 was detected by flow cytometry.
Results: ZZR-medicated sera with volume fraction of 8%, 16% and 32% of medium dose were confirmed as the experimental sera. Compared with the untreated group, NF-κB activities of the ZZR-medicated serum groups (ZZR-medicated serum with volume fraction of 8%, 16% and 32% of medium dose) were obviously down-regulated (P<0.01), which had a negative correlation with the concentrations. After interfering HCT-8/VCR with ZZR-medicated serum of different concentrations for 24 h, P-gp in HCT-8/VCR transmitted gradually from cell membrane to cytoplasm and nuclei. Nuclei became pyknotic and cracking. Compared with the untreated group, the mean fluorescence intensities of ZZR-medicated serum groups declined with concentration gradients (P<0.01). The efflux of intracellular rhodamine 123 decreased, the wave crest shifted to right, and the intracellular fluorescence intensity strengthened (P<0.01).
Conclusion: ZZR-medicated sera of experimental concentrations down-regulate activity of NF-κB and expression and function of P-gp in human colorectal cancer multidrug-resistant cell line HCT-8/VCR and the effect is related to the concentrations.

Key words: colorectal neoplasms, P-glycoprotein, NF-kappa B, compound (traditional Chinese drugs)

Table 1

Sensitivity of HCT-8 and HCT-8/VCR cells to different chemotherapeutic drugs (x±s)"

Drug n IC50 (μg/mL)
HCT-8 HCT-8/VCR Resistance index of HCT-8/VCR (%)
VCR 3 0.14±0.02 9.55±0.23** 70.74
5-FU 3 3.19±0.34 65.71±0.26** 20.60
CDDP 3 0.74±0.04 4.13±0.16** 5.58
EP 3 0.80±0.07 3.79±0.12** 4.74

Table 2

Inhibition rates of HCT-8/VCR cell line treated with different concentrations of medicated serum"

Group n Inhibition rate at different concentrations of medicated serum
2% 4% 8% 16% 32% 64%
Blank serum 3 1.24±0.31 3.56±0.01 2.14±0.02 2.77±0.05 3.43±0.05 3.35±0.08
Low-dose medicated serum 3 0 7.42±0.03 12.91±1.68△△ 11.19±1.15△△ 10.59±2.07△△ 41.10±1.02△△
Medium-dose medicated serum 3 1.61±1.12 11.03±1.31 20.11±1.83△△ 23.70±1.61△△ 24.75±2.86△△ 43.89±3.28△△
High-dose medicated serum 3 1.43±0.07 9.45±0.04 16.24±1.04△△ 21.32±1.59△△ 30.29±3.46△△ 57.71±3.69△△

Table 3

NF-κB activity in HCT-8/VCR cell line treated with medicated serum for 24 h (x±s)"

Group n Absorbance value at 450 nm
HCT-8 3 3.534±0.120*
HCT-8/VCR 3 3.863±0.055
Blank serum 3 3.923±0.125
8% Medicated serum 3 3.457±0.091△△
16% Medicated serum 3 2.946±0.066△△
32% Medicated serum 3 2.868±0.079△△
BAY11-7082 3 2.730±0.042△△

Figure 1

Distribution of P-gp in HCT-8 and HCT-8/VCR cell lines treated with different concentrations of medicated serum (Laser scanning confocal microscopy, ×2 400) A: HCT-8; B: HCT-8/VCR; C: Blank serum; D: 8% Medicated serum; E: 16% Medicated serum; F: 32% Medicated serum; G: BAY11-7082. P-gp: P-glycoprotein."

Table 4

Mean fluorescence intensity of P-gp in cells treated with different concentrations of medicated serum"

Group n Mean flourence intensity
HCT-8 3 33.175±3.735
HCT-8/VCR 3 133.015±6.548
Blank serum 3 134.090±10.400
8% Medicated serum 3 43.148±3.236△△
16% Medicated serum 3 25.840±2.640△△
32% Medicated serum 3 23.448±3.302△△
BAY11-7082 3 8.625±4.406△△

Table 5

Efflux function of P-gp in cells treated with different concentrations of medicated serum"

Group n Mean flourence intensity
HCT-8 3 100.00
HCT-8/VCR 3 24.82±12.07
Blank serum 3 26.31±1.04
8% Medicated serum 3 186.76±40.42△△
16% Medicated serum 3 245.78±10.09△△
32% Medicated serum 3 282.31±77.53△△
BAY11-7082 3 321.69±20.36△△

Figure 2

Efflux function of P-gp in cells treated with different concentrations of medicated serum A: HCT-8; B: HCT-8/VCR; C: Blank serum; D: 8% Medicated serum; E: 16% Medicated serum; F: 32% Medicated serum; G: BAY11-7082. P-gp: P-glycoprotein."

[1] Luo XQ, Yang GW . Research progress on traditional Chinese medicine treatment of gastric cancer. Yi Xue Zong Shu. 2009; 15(18): 2835-2838. Chinese with abstract in English.
罗晓琴, 杨国旺 . 中医药治疗胃癌的研究进展.医学综述. 2009; 15(18):2835-2838.
[2] Xu SY, Bian RL, Chen X. Methodology of pharmacological experiment. Beijing: People’s Medical Publishing House. 1982: 202-204. Chinese.
徐叔云, 卞如濂, 陈修 . 药理实验方法学.北京: 人民卫生出版社. 1982: 202-204.
[3] Feller N, Kuiper CM, Lankelma J, Ruhdal JK, Scheper RJ, Pinedo HM, Broxterman HJ . Functional detection of MDR1/P170 and MRP/P19〇-mediated multidrug resistance in tumour cells by flow cytometry[J]. Br Cancer. 1995; 72(3) : 543-549.
doi: 10.1038/bjc.1995.371
[4] Haimeur A, Conseil G, Deeley RG, Cole SP . The MRP-related and BCRP/ABCG2 multidrug resistance proteins : biology, substrate specificity and regulation. Curr Drug Metab. 2005; 5(1):21-53.
[5] Yeh JJ, Hsu NY, Hsu WH, Tsai CH, Lin CC, Liang JA . Comparison of chemotherapy response with P- glycoprotein, multidrug resistance-related protein-1, and lung resistance-related protein expression in untreated small cell lung cancer. Lung. 2005 ; 183(3):177-183.
doi: 10.1007/s00408-004-2532-1
[6] Berney DM, Shamash J, Gaffney J, Jordan S, Oliver RT . DNA topoisomerase I and E expression in drug resistant germ cell tumours[J]. Br Cancer. 2002 ; 87(6):624-629.
doi: 10.1038/sj.bjc.6600472 pmid: 2364243
[7] Notarbartolo M, Cervello M, Dusonchet L, Cusimano A, D’Alessandro N . Resistance to diverse apoptotic triggers in multidrug resistant HL60 cells and its possible relationship to the expression of P-glycoprotein, Fas and of the novel anti-apoptosis factors IAP (inhibitory of apoptosis proteins). Cancer Lett. 2002 ; 180(1) : 91-101.
doi: 10.1016/S0304-3835(01)00834-5
[8] Queiroz KC, Ruela-de-Sousa RR, Fuhler GM, Aberson HL, Ferreira CV, Peppelenbosch MP, Spek CA . Hedgehog signaling maintains chemoresistance in myeloid leukemic cells. Oncogene. 2010; 29(48):6314-6322.
doi: 10.1038/onc.2010.375 pmid: 20802532
[9] Hayden MS, Ghosh S . Shared principles in NF-icB signaling. Cell. 2008; 132(3):344-362.
doi: 10.1016/j.cell.2008.01.020
[10] Bentires-Alj M, Barbu V, Fillet M, Chariot A, Relic B, Jacobs N, Gielen J, Merville MP, Bours V . NF-icB transcription factor induces drug resistance through MDR1 expression in cancer cells. Oncogene. 2003; 22(1):90-97.
doi: 10.1038/sj.onc.1206056
[11] Bourguignon LY, Xia W, Wong G . Hyaluronan-mediated CD44 interaction with p300 and SIRT1 regulates p-catenin signaling and NFicfrspecific transcription activity leading to MDR1 and Bcl-xL gene expression and chemoresistance in breast tumor cells.[J] Biol Chem. 2009 ; 284(5):2657-2671.
doi: 10.1074/jbc.M806708200
[12] Zhang JK, Wang ZY, Ding DP, Xin Y . Effect of Astragulus injection on chemosensitivity of drug-resistant hepatic carcinoma Bel/Fu cell line. Zhongguo Zhong Xi Yi Jie He Wai Ke Za Zhi. 2008; 14(4): 398-402. Chinese with abstract in English.
doi: 10.3969/j.issn.1007-6948.2008.04.039
张隽开, 王忠裕, 丁大朋, 辛毅 . 黄芪对肝癌耐药细胞株Bel/Fu化疗敏感性的影响.中国中西医结合外科杂志. 2008; 14(4):398-402.
doi: 10.3969/j.issn.1007-6948.2008.04.039
[1] Li Tao, Ying-jie Zhu , Xian-mei Lu , Ying Gu , Ai-guang Zhao , Jian Zheng, Chuan-gang Fu, Jin-kun Yang. Clinical study on survival benefit for elderly patients with resected stage I or E colorectal cancer based on traditional Chinese medicine syndrome differentiation and treatment. Journal of Chinese Integrative Medicine, 2010, 8(12): 1159-1164.
[2] Qing Lin , Jian Zheng, Si Ai, Li-xing Lin, Yu-xin Chen, Fang Wang, Ling Zhu . Mechanism of combined treatment with Shenkangling Decoction and prednisone in rats with adriamycin-induced nephropathy. Journal of Chinese Integrative Medicine, 2009, 7(7): 661-666.
[3] Ju-yong Wang , Ye Jin, Zhen-ye Xu, Zhan Zheng. Effects of Feiyanning Decoction on gene expression of nuclear factor-κB activated by tumor necrosis factor-α in lung adenocarcinoma cell line. Journal of Chinese Integrative Medicine, 2009, 7(3): 249-254.
[4] Gong-jian Dai, Hei-ying Jin, Yi-jiang Ding, Jian-guo Xia, Xiu-fang Liu, Fei Liu, Xuan-zhong Tan, Jian-xiang Geng. Anticancer effects of tea polyphenols on colorectal cancer with microsatellite instability in nude mice. Journal of Chinese Integrative Medicine, 2008, 6(12): 1263-1266.
Viewed
Full text


Abstract

Cited

  Shared   
  Discussed   
[1] Dong Yang, Yong-ping Du, Qing Shen, Wei Chen, Yan Yu, Guang-lei Chen. Expression of alpha-smooth muscle actin in renal tubulointerstitium in patients with kidney collateral stasis. Journal of Chinese Integrative Medicine, 2008, 6(1): 41-44
[2] A-gao Zhou, Yong Zhang, Gang Kui, De-Yun Kong, Hai-liang Ge, Qiu-hua Ren, Jia-rong Dong, Sheng Hong, Xu-ming Mao, Yin Wang, Hui-zheng Zhang, Shu-jun Wang. Influence of traditional Chinese compound recipes with different efficacy on body weight, tumor weight and immune function in H22 cancer-bearing mice. Journal of Chinese Integrative Medicine, 2008, 6(1): 77-82
[3] 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
[4] Bo Wang , Wei Yan , Li-hui Hou, Xiao-ke Wu. Disorder of Tiangui (kidney essence) and reproductive dysfunction in patients with polycystic ovary syndrome. Journal of Chinese Integrative Medicine, 2010, 8(11): 1018-1022
[5] Ya-hui Liang , Ping Li, Jing-xia Zhao, Xin Liu, Qi-fu Huang. Acetyl-11-keto-beta-boswellic acid and arsenic trioxide regulate the productions and activities of matrix metalloproteinases in human skin fibroblasts and human leukemia cell line THP-1. Journal of Chinese Integrative Medicine, 2010, 8(11): 1060-1069
[6] Pedro Saganha João, Doenitz Christoph, Greten Tobias, Efferth Thomas, J. Greten Henry. Qigong therapy for physiotherapists suffering from burnout: a preliminary study. Journal of Chinese Integrative Medicine, 2012, 10(11): 1233-1239
[7] Xiang-ying Mao , Qin Bian , Zi-yin Shen. Analysis of the osteogenetic effects exerted on mesenchymal stem cell strain C3H10T1/2 by icariin via MAPK signaling pathway in vitro. Journal of Chinese Integrative Medicine, 2012, 10(11): 1272-1278
[8] Xiao Zhang , Bing-bing Ning , Shuang Ren , Li-jun Zhang , Wen-meng Zhang , Jia-mei Chen , Gao-feng Chen , Hua Zhang , Yong-ping Mu , Ping Liu. Effects of Chinese herbal medicine Xiaopi Pill in preventing rats from dimethylnitrosamine-induced liver fibrosis. Journal of Chinese Integrative Medicine, 2012, 10(11): 1286-1292
[9] Ji Chen, Qin Pan, Li-shuang Ye, Jia-ling Huang. English translation of cultural aspects of the titles of traditional Chinese medicine classics based on the skopos theory. Journal of Chinese Integrative Medicine, 2012, 10(11): 1316-1320
[10] Peng-fei Gao, Kenji Watanabe. Introduction of the World Health Organization project of the International Classification of Traditional Medicine. Journal of Chinese Integrative Medicine, 2011, 9(11): 1161-1164