Paclitaxel and cisplatin inhibit the proliferation of nasopharyngeal cancer stem cells and promote apoptosis via the Wnt/beta-catenin pathway

doi:10.3969/j.issn.2095-4344.2016.41.006

Abstract

Abstract:

BACKGROUND: Cancer stem cells have self-renewal ability and can differentiate into new tumors. Cancer stem cells are the source of tumor formation and recurrence, and they can make tumors insensitive to radiotherapy and chemotherapy.
OBJECTIVE: To explore the effect of paclitaxel plus cisplatin on the proliferation and apoptosis of nasopharyngeal cancer stem cells (NPCSCs) and involved signal pathways.
METHODS: NPCSCs were sorted by immunomagneticbeads and were treated with paclitaxel, cisplatin or their combination. The expression of caspase-3, activated caspase-3 and Bcl-2, which are related to apoptosis, was determined by western blot. The expression of β-catenin and its downstream proto-oncogene, c-myc, was also determined by western blot. The activity of the Wnt/β-catenin pathway was inhibited by knocking down β-catenin expression or β-catenin inhibitor XAV939. Proliferation and apoptosis of NPCSCs were detected by MTT and flow cytometry, respectively.
RESULTS AND CONCLUSION: Either paclitaxel or cisplatin could inhibit proliferation and induce apoptosis of NPCSCs. The expression of apoptosis marker, activated caspase-3, was increased and the expression of the inhibitor of apoptosis, Bcl-2, was declined. Combined use of paclitaxel and cisplatin had synergistic effect when used together. Either paclitaxel or cisplatin could inhibit the expression of β-catenin and c-myc, suppressed the proliferation and induced the apoptosis of NPCSCs by inhibiting the activity of Wnt/β-catenin pathway. These results indicate that the combined use of paclitaxel and cisplatin may inhibit the proliferation of NPCSCs and promote apoptosis via the Wnt/β-catenin pathway.

中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程

Key words: Stem Cells, Drug Therapy, Signal Transduction, Cell Proliferation, Apoptosis, Tissue Engineering

CLC Number:

R318

Liu Yong-gang, Yang Rong-song, Wu Hong-fang, Zhang Bao-chao . Paclitaxel and cisplatin inhibit the proliferation of nasopharyngeal cancer stem cells and promote apoptosis via the Wnt/beta-catenin pathway[J]. Chinese Journal of Tissue Engineering Research, 2016, 20(41): 6118-6124.

Figures/Tables 2

2.1 不同质量浓度紫杉醇和顺铂对鼻咽癌干细胞增殖的影响为了摸索合适的药物浓度用于后续实验，使用不同浓度的紫杉醇或顺铂处理鼻咽癌干细胞，MTT法检测细胞的增殖情况。结果如图1，紫杉醇和顺铂均能抑制体外培养的鼻咽癌干细胞的增殖，且呈剂量依赖性。综合考虑各方面因素，选择4 mg/L的紫杉醇和8 mg/L顺铂用于后续实验。 2.2 紫杉醇联合顺铂对鼻咽癌干细胞增殖和凋亡的影响为了研究紫杉醇联合顺铂对鼻咽癌干细胞凋亡的影响，分别用紫杉醇、顺铂、紫杉醇联合顺铂处理细胞，MTT检测细胞增殖率，流式细胞仪检测细胞凋亡情况，Western blot检测凋亡相关蛋白Bcl-2、Caspase-3和活化caspase-3表达。结果表明，紫杉醇或顺铂处理细胞后细胞增殖受到抑制，细胞凋亡率上升，凋亡标志蛋白活化caspase-3表达上升，凋亡抑制蛋白Bcl-2表达下降，联合用药较单一用药抑制增殖和促凋亡效果更好(图2)。这些结果表明紫杉醇和顺铂均可抑制鼻咽癌干细胞增殖，促进其凋亡，并且两种药物具有协同作用。 2.3 紫杉醇联合顺铂作用细胞后β-catenin及下游c-myc蛋白的表达为了研究紫杉醇联合顺铂对鼻咽癌干细胞Wnt/β-catenin通路的影响，用紫杉醇和顺铂处理鼻咽癌干细胞后Western blot检测β-catenin及下游原癌基因c-myc的表达。如图3所示，紫杉醇或顺铂对β-catenin及c-myc的表达均有不同程度的抑制作用，2种药物联合处理对Wnt/β-catenin通路的抑制作用更强，表明紫杉醇和顺铂可协同抑制Wnt/β-catenin通路活性。 2.4 使用RNA干扰或抑制剂抑制Wnt/β-catenin通路活性后鼻咽癌干细胞增殖和凋亡的变化为了探索紫杉醇和顺铂抑制鼻咽癌干细胞增殖和促进凋亡是否通过Wnt/β-catenin通路，使用RNA干扰敲降β-catenin表达或使用抑制剂XAV939抑制Wnt/β-catenin通路活性后检测鼻咽癌干细胞增殖和凋亡的变化。图4结果发现，敲降β-catenin表达或抑制剂抑制Wnt/β-catenin通路活性后鼻咽癌干细胞增殖受到抑制，凋亡增加，Bcl-2表达下降，活化caspase-3表达上升，表明Wnt/β-catenin通路活性的变化对鼻咽癌干细胞的增殖和凋亡有作用。"

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