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纸质出版:2016
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泛素水解酶22 siRNA对宫颈癌CD133+ 宫颈癌干细胞增殖和侵袭的影响[J]. 中山大学学报(医学科学版), 2016,37(1).
Effect of siRNA USP22 on Proliferation and Invasion of CD133+ Cervical Cancer Stem Cells[J]. Journal of Sun Yat-sen University (Medical Sciences), 2016, 37(1).
摘 要: 【目的】 采用USP22 siRNA对宫颈癌CD133+CaSki宫颈癌干细胞进行基因治疗,观察其对USP22含量和对增殖侵袭能力的影响,并对其分子生物学机制进行初步探讨。【方法】 流式分选获得CD133+ CaSki宫颈癌干细胞,分为USP22组、阴性对照(NC)组、空白对照组(MOCK)组。采用western blot法比较各组细胞USP22蛋白的含量,MTT法检测3组细胞的增殖能力,Transwell法检测3组细胞的侵袭能力,荧光素酶报告基因实验检测3组细胞Wnt/β-catenin信号通路的活性。【结果】 采用流式细胞术分选CD133阳性细胞后CaSki细胞CDl33阳性率为 (96.87 ± 5.31)%,显示流式分选宫颈癌干细胞成功(P < 0.05)。Western blot检测结果显示,USP22组CD133+ CaSki细胞中USP22蛋白表达量明显降低。MTT结果显示USP22组细胞增殖能力明显减弱,差异具有统计学意义(P < 0.05)。Transwell实验结果显示USP22组CD133+ CaSki细胞侵袭能力明显受到抑制,差异具有统计学意义(P < 0.05)。荧光素酶报告基因实验结果显示,USP22组Wnt/β-catenin信号通路的转录活性明显降低,差异具有统计学意义(P < 0.05)。【结论】 USP22 siRNA有降低CD133+ CaSki宫颈癌干细胞内Wnt/β-catenin信号通路活性的作用,进而可以实现对宫颈癌干细胞增殖和侵袭功能的抑制,该治疗方法有望成为宫颈癌基因治疗的新手段。
Abstract: 【Objective】 CD133+CaSki cervical cancer stem cells transfected by USP22 siRNA, observe the content of USP22 and its effect on proliferation and invasion, and its molecular mechanisms are discussed. 【Methods】 Flow sorting obtain CD133+CaSki cervical cancer stem cells were divided into USP22 group, negative control (NC) group and blank control (MOCK) group. Western blot was used to compare the content of USP22 protein in each group. The proliferation ability of the 3 groups of cells was detected by MTT assay. Transwell method was used to detect the invasion ability of the 3 groups of cells. Luciferase reporter gene assay was used to detect the activity of Wnt/ beta -catenin signaling pathway in 3 groups of cells. 【Results】 The positive rates of CDl33 cells after flow sorting were (96.87 ± 5.31)%, elected cervical cancer stem cells successfully (P < 0.05). The expression of USP22 protein in USP22 group was significantly lower than that in MOCK group. MTT results showed that the proliferation ability of USP22 group was significantly decreased, the difference was statistically significant (P < 0.05). The number of invasion cells in USP22 group was significantly lower than that in MOCK group, the difference was statistically significant (P < 0.05). The transcriptional activity of Wnt/β-catenin signaling pathway in USP22 group was significantly lower than that in MOCK group, the difference was statistically significant (P < 0.05). 【Conclusion】 USP22 siRNA can inhibit proliferation and invasion of CD133+CaSki cervical cancer stem cells by down regulating the transcriptional activity of the β-catenin signaling pathway. This treatment is expected to become the new means of gene therapy in cervical carcinoma.
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