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Journal of Chinese Integrative Medicine ›› 2009, Vol. 7 ›› Issue (10): 952-957.doi: 10.3736/jcim20091008

• Original Experimental Research • Previous Articles     Next Articles

Resveratrol increases sensitivity of CNE2 cells to chemotherapeutic drugs under hypoxia

Fang Quana(),Shao-qiang Zhanga,Yan-xia Baia,Xiao-bao Yaoa,Hong-hui Lia,LiangYub,Cheng-en Panb   

  1. a Department of ENT & Head and Neck Surgery, the First Affiliated Hospital, Xi’an Jiao Tong University, Xi’an 710061, Shaanxi Province, China;
    b Department of Hepatobiliary Surgery, the First Affiliated Hospital, Xi’an Jiao Tong University, Xi’an 710061,Shaanxi Province, China;
  • Received:2009-04-27 Accepted:2009-07-06 Online:2009-10-20 Published:2009-10-15
  • Contact: Fang Quan


To explore the sensitization effects of resveratrol on CNE2 nasopharyngeal carcinoma cell line with hypoxia-induced chemotherapy resistance and the potential mechanism.

Human CNE2 nasopharyngeal carcinoma cell line was cultured under hypoxic conditions (37 ℃, 5% CO2, 2% O2) in vitro. The cultured cells were treated with different concentrations of resveratrol for 48 h. Reversal fold (RF) of reseratrol to chemotherapeutic drugs in CNE2 cells was measured by methyl thiazolyl tetrazolium (MTT) assay. Apoptotic rate of CNE2 cells was observed by flow cytometry. Reverse transcription-polymerase chain reaction (RT-PCR) method and Western blotting were used to investigate the expressions of multidrug resistance gene 1 (mdr1), multidrug resistance-associated protein 1 (MRP1) and hypoxia inducible factor 1α (HIF-1α) in CNE2 cells.

Resveratrol combined with chemotherapeutics produced a synergistic effect. The RF of 200 μmol/L resveratrol to paclitaxel was 2.58. Combined with paclitaxel, 25, 50, 100 and 200 μmol/L of resveratrol increased the apoptotic rate of CNE2 cells from (22.14±1.09)% to (23.24±1.37)%, (27.57±2.01)%, and (30.36±2.31)%, respectively. Resveratrol could down-regulate the expressions of HIF-1α, mdr1 and MRP1 significantly. After being treated with resveratrol at different concentrations separately, the expressions of HIF-1α, mdr1 and MRP1 in CNE2 cells decreased significantly as compared with paclitaxel alone or paclitaxel plus verapamil (P<0.01).

Resveratrol can enhance the sensitivity of CNE2 cells to chemotherapeutic drugs under hypoxia. The potential mechanism is partly attributed to inhibiting the gene expressions of HIF-1α, mdr1 and MRP1.

Key words: Resveratrol, Chemotherapy sensitization, Hypoxia, Nasopharyngeal carcinoma

Table 1

Effects of resveratrol in modulating the sensitivity of CNE2 cells to cytotoxic drugs under hypoxia ($\bar{x}$±s, mol/L)"

Group n Paclitaxel Cisplatin 5-Fluorouracil
IC50 (×10–8) RF IC50 (×10–6) RF IC50 (×10–5) RF
Hypoxia 4 3.40±0.52 1.48±0.23 9.39±0.81
Hypoxia plus 25 μmol/L Res 4 2.98±0.45 1.14 1.25±0.38 1.18 8.77±0.30 1.07
Hypoxia plus 50 μmol/L Res 4 2.56±0.71 1.33 1.11±0.19 1.34 7.57±0.16 1.24
Hypoxia plus 100 μmol/L Res 4 1.86±0.66 1.89 0.97±0.03 1.52 6.75±0.27 1.39
Hypoxia plus 200 μmol/L Res 4 1.32±0.50 2.58 0.85±0.04 1.75 6.02±0.35 1.56

Table 2

Apoptosis of CNE2 cells induced by paclitaxel plus resverarol under hypoxia observed by flow cytometer ($\bar{x}$±s, %)"

Group n Rate of apoptosis
Paclitaxel 5 20.70±1.11
Paclitaxel plus 10 μmol/L verapamil 5 32.49±2.21**
Paclitaxel plus 25 μmol/L Res 5 22.14±1.09**△△▲▲
Paclitaxel plus 50 μmol/L Res 5 23.24±1.37**△△▲▲
Paclitaxel plus 100 μmol/L Res 5 27.57±2.01**△△▲
Paclitaxel plus 200 μmol/L Res 5 30.36±2.31**

Figure 1

Apoptosis of CNE2 cells induced by paclitaxel plus resverarol under hypoxia observed by flow cytometry CNE2 cells were incubated with 8 nmol/L paclitaxel in the presence of various concentrations of resveratrol or 10 μmol/L verapamil for 48 h under hypoxia. The rate of apoptosis was measured by using flow cytometry. A: Paclitaxel; B: Paclitaxel plus verapamil; C-F: Paclitaxel plus 25, 50, 100 and 200 μmol/L of resveratrol, respectively."

Table 3

Effects of resveratrol on expressions of mdr1, MPR1 and HIF-1α mRNAs in CNE2 cells ($\bar{x}$±s, %)"

Group n MRP1 mdr1 HIF-1α
Paclitaxel 5 0.79±0.05 0.42±0.04 0.98±0.04
Paclitaxel plus 10 μmol/L verapamil 5 0.32±0.03** 0.13±0.01** 0.49±0.01**
Paclitaxel plus 25 μmol/L Res 5 0.71±0.02**△△▲▲ 0.31±0.03**△△▲▲ 0.91±0.03**△△▲▲
Paclitaxel plus 50 μmol/L Res 5 0.67±0.04**△△▲▲ 0.27±0.03**△△▲▲ 0.85±0.03**△△▲▲
Paclitaxel plus 100 μmol/L Res 5 0.55±0.03**△△▲▲ 0.19±0.04**△△ 0.73±0.04**△△▲▲
Paclitaxel plus 200 μmol/L Res 5 0.42±0.02**△△ 0.15±0.02** 0.61±0.02**

Figure 2

Expressions of mdr1, MPR1 and HIF-1α mRNAsin CNE2 cells induced by paclitaxel plus resverarol underhypoxia observed by RT-PCR"

Figure 3

Expressions of P-gp, MRP1 and HIF-1α proteinsin CNE2 cells induced by paclitaxel plus resveratrol underhypoxia observed by Western blotting"

Table 4

Effects of resveratrol on expressions of P-gp, MRP1 and HIF-1α proteins in CNE2 cells ($\bar{x}$±s)"

Group n MRP1 P-gp HIF-1α
Paclitaxel 5 0.61±0.03 1.20±0.02 1.27±0.04
Paclitaxel plus 10 μmol/L verapamil 5 0.11±0.01* 0.75±0.03** 0.62±0.03**
Paclitaxel plus 25 μmol/L Res 5 0.62±0.01△△▲▲ 1.15±0.04*△△▲▲ 1.15±0.20**△△▲▲
Paclitaxel plus 50 μmol/L Res 5 0.57±0.06△△▲▲ 1.10±0.03**△△▲▲ 1.06±0.03**△△▲▲
Paclitaxel plus 100 μmol/L Res 5 0.41±0.03**△△▲▲ 0.91±0.40**△△▲▲ 0.91±0.04**△△▲▲
Paclitaxel plus 200 μmol/L Res 5 0.32±0.05**△△ 0.82±0.02**△△ 0.79±0.07**△△
[1] Quan F, Pan C, Ma Q, Zhang S, Yan L . Reversal effect of resveratrol on multidrug resistance in KBv200 cell line[J]. Biomed Pharmacother, 2008,62(9):622-629
doi: 10.1016/j.biopha.2008.07.089 pmid: 18804944
[2] Xu S, Luo L, Zhu LQ, Li Z, Qiu LL, Chen Q, Xu CF . Reversal effect of 4'-methylether-scutellarein on multidrug resistance of human choriocarcinoma JAR/VP16 cell line[J]. Sheng Wu Hua Xue Yu Sheng Wu Wu Li Jin Zhan, 2006,33(11):1061-1073
徐珊, 罗莉, 朱利群, 李卓, 仇黎丽, 陈琪, 徐昌芬 . 4'-甲醚-黄芩素对绒癌耐药细胞株多药耐药性的逆转作用研究[J]. 生物化学与生物物理进展, 2006,33(11):1061-1073
[3] Shannon AM, Bouchier-Hayes DJ, Condron CM, Toomey D . Tumor hypoxia, chemotherapeutic resistance and hypoxia-related therapies[J]. Cancer Treat Rev, 2003,29(4):297-307
doi: 10.1016/S0305-7372(03)00003-3
[4] El-Saggan AH, Dovinova I, Sulova Z, Barancik M, Hunakova L, Breier A, Uhrik B . Hypoxia increases cell death in multidrug-resistant leukemia cells. Differences in viability and ultrastructure between sensitive and multidrug-resistant L1210 mouse leukemic cells under hypoxia[J]. Gen Physiol Biophys, 2003,22(2):265-273
[5] Vaupel P . The role of hypoxia-induced factors in tumor progression[J]. Oncologist, 2004,9(Suppl 5):10-17
doi: 10.1634/theoncologist.9-90005-10 pmid: 15591418
[6] Munoz-Martinez F, Lu P, Cortes-Selva F, Perez-Victoria JM, Jimenez IA, Ravelo AG, Sharom FJ, Gamarro F, Castanvs S . Celastraceae sesquiterpenes as a new class of modulators that bind specifically to human P-glycoprotein and reverse cellular multi-drug resistance[J]. Cancer Res, 2004,64(19):7130-7138
doi: 10.1158/0008-5472.CAN-04-1005
[7] Yang R , McBride A, Hou Y X, Goldberg A, Chang XB. Nucleotide dissociation from NBD1 promotes solute transport by MRP1[J]. Biochim Biophys Acta, 2005,1668(2):248-261
doi: 10.1016/j.bbamem.2004.12.013 pmid: 15737336
[8] Lee JW, Bae SH, Jeong JW, Kim SH, Kim KW . Hypoxia-inducible factor(HIF-1) alpha: its protein stability and biological functions[J]. Exp Mol Med, 2004,36(1):1-12
doi: 10.1038/emm.2004.1
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