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. 2014 Sep 15;7(10):6784-91.
eCollection 2014.

MiR-634 sensitizes nasopharyngeal carcinoma cells to paclitaxel and inhibits cell growth both in vitro and in vivo

Xiaowei Peng  1 , Peiguo Cao  2 , Dong He  3 , Shuang Han  2 , Jianda Zhou  4 , Guolin Tan  5 , Wei Li  5 , Fenghui Yu  1 , Jianjun Yu  1 , Zan Li  1 , Ke Cao  2
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MiR-634 sensitizes nasopharyngeal carcinoma cells to paclitaxel and inhibits cell growth both in vitro and in vivo

V体育ios版 - Xiaowei Peng et al. Int J Clin Exp Pathol. .

Abstract

Resistance to chemotherapy is one of the key causal factors in cancer death and increasing evidence has revealed that microRNAs (miRNAs) are involved in chemoresistance in many kinds of human cancers VSports手机版. Paclitaxel has been used for treatment of advanced nasopharyngeal carcinoma (NPC); however, treatment failure often occurs due to development of acquired paclitaxel resistance. In this study, based on miRNA microarray screening and qRT-PCR validation, we found six differentially expressed miRNAs in our induced paclitaxel-resistant NPC CNE-1/Taxol cells. Furthermore, we clarified the role of miR-634, most significantly downregulated in the paclitaxel-resistant CNE-1/Taxol, in regulating the paclitaxel sensitivity in NPC cells. We restored miR-634 expression in the CNE-1/Taxol cells by lentivirus infection, and found restoration of miR-634 re-sensitized the CNE-1/Taxol cells to paclitaxel in vitro by MTT assay and colony formation assay. In xenograft mouse model, we found that miR-634 inhibited tumor growth and enhanced paclitaxel sensitivity. Thus, our findings provide important information for the development of targeted gene therapy for reversing paclitaxel resistance in NPC. .

Keywords: Microarray; nasopharyngeal carcinoma; paclitaxel resistance; targeted therapy V体育安卓版. .

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Figure 1
Figure 1
Screening and validation of miRNAs associated with paclitaxel resistance in CNE-1 cells. A. Hierarchical clustering analysis of 13 differentially expressed miRNAs detected by the miRNA microarray analysis in the paclitaxol-resistant CNE-1/Taxol cell sublines (n = 3, N2, N4, N6) compared to the parental CNE-1 cells (n = 3, Q1, Q3, Q5). The heat map diagram shows the results of the two-way hierarchical clustering analysis of miRNA expression levels and the cell lines. Each row represents a miRNA and each column represents a cell line. The miRNA-clustering tree is shown on the left and the cell line-clustering tree is at the top. The color scale shown at the top illustrates the relative expression level of a miRNA in a certain slide. A red color represents a high relative expression level and a green color represents a low relative expression level. B. Six differentially expressed miRNAs were validated by qRT-PCR in the paclitaxol-resistant CNE-1/Taxol cell sublines. There were five significantly downregulated (miR-634, miR-4478, miR-3198, miR-100 and miR-4653) and one significantly upregulated (miR-802) miRNAs consistent with the microarray results.
Figure 2
Figure 2
MiR-634 sensitizes nasopharyngeal carcinoma cells to paclitaxel in vitro. A. The CNE-1/Taxol cells were infected with Lv-miR-634 or Lv-NC, after being infected for 6 days, qRT-PCR was performed to determinate expression levels of miR-634 in stably infected CNE-1/Taxol cells, untreated groups were taken as the control. B. The impact of miR-634 on drug sensitivity at different paclitaxel doses (0, 2, 4, 6, 8 and 10 ng/ml) was determined by MTT assay. C. The CNE-1/Taxol cells were treated with a final concentration of 10 ng/ml paclitaxel for 24 h, and the impact of miR-634 on drug sensitivity was determined by colony formation assay. *P < 0.05.
Figure 3
Figure 3
MiR-634 inhibits tumor growth and sensitizes nasopharyngeal carcinoma cells to paclitaxel in vivo. BALB/C nude mice were subcutaneously inoculated with CNE-1/Taxol cells with overexpressed miR-634 (Lv-miR-634, n = 5) and negative controls (Lv-miR-NC, n = 5), respectively. After 61 days, paclitaxel (10 mg kg-1) was then intravenously injected into mice once a day for five days. A. Tumor volume (mm3) was calculated every 4 days. B. Two days after complete paclitaxel treatments, all mice were euthanized and the tumors were excised and imaged under a light microscope. C. qRT-PCR analysis showed that miR-634 was upregulated in transplanted tumour tissues inoculated by miR-634-overexpressing CNE-1/Taxol cells. *P < 0.05.
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