Epigenetic modification regulation of Wnt pathway inhibitors to study the mechanism of polyphyllinⅦ against colorectal cancer

doi:10.3877/cma.j.issn.2095-3224.2019.06.006

Abstract

Abstract:

Objective

Inquiry PolyphyllinⅦ resistance mechanism of colorectal cancer from Wnt pathways inhibitory factor the epigeNETsic modifications regulation.

Methods

The experiment is divided into three groups: blank control group, model group and colorectal cancer polyphyllinⅦ treatment group. TUNEL assay was used to detect apoptosis in mice. TOPglow/FOPglow TCF kit was used to detect activity of Wnt signal transduction pathway. Sandwich ELISA was used to detect expression of caspase-3, caspase-8, caspase-9, Bax, bcl-2, cdk-4, cdk-6, Cyclin D1 and p21. The expression of stem cell related genes OCT4, SOX2, c-myc and TCF4 was detected by real-time fluorescence quantitative PCR, and the binding of lgG, beta-catenin and H3k9Ac to mmp-9 promoter was detected.

Results

Compared with the control group, TUNEL immunohistochemical analysis showed that PPLVII significantly induced apoptosis of tumor cells. After PPLVII treatment, cleaved caspase-3, cleaved caspase-8, cleaved caspase-9 and Bax expression were increased, while Bcl-2 expression was decreased, the difference was significant (P<0.05). The expression of Cyclin D1, CDK-4 and CDK-6 decreased, while the expression of p21 was up-regulated, the difference was significant (P<0.01). The TOP/FOP ratio in the treatment group was decreased after PPLVII intervention. Betacatenin was detected by western blotting, The results showed that the gray level of PPLVII intervention decreased, the difference was significant (P<0.05). The expressions of OCT4, SOX2, c-Myc and TCF4 related genes were down-regulated after PPLVII intervention, the difference was significant (P<0.05).

Conclusion

Treatment by reducing PPLVII Wnt pathways of inhibiting factor expression in colorectal cancer have certain improvement and treatment effect.

Key words: Colorectal neoplasms, Wnt pathways inhibitory factor, Polyphyllin Ⅶ

Mingwang Chen, Shuying Gao, Haiguang Liu, Qingzhu Tang, Hongbiao Luo. Epigenetic modification regulation of Wnt pathway inhibitors to study the mechanism of polyphyllinⅦ against colorectal cancer[J]. Chinese Journal of Colorectal Diseases(Electronic Edition), 2019, 08(06): 574-579.

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Figures/Tables 5

References 20

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组别	例数	cleaved-caspase-3	cleaved-caspase-8	cleaved-caspase-9	Bax	Bcl-2
对照组	10	0.75±0.02	0.53±0.04	0.39±0.01	0.26±0.02	0.29±0.04
模型组	10	0.19±0.03	0.11±0.03	0.12±0.02	0.15±0.02	0.81±0.02
治疗组	10	0.61±0.02	0.48±0.03	0.32±0.01	0.25±0.03	0.38±0.03
F值	?	22.962	19.281	16.035	16.859	15.364
P值	?	0.001	0.001	0.005	0.007	0.010

组别	例数	cleaved-caspase-3	cleaved-caspase-8	cleaved-caspase-9	Bax	Bcl-2
对照组	10	0.75±0.02	0.53±0.04	0.39±0.01	0.26±0.02	0.29±0.04
模型组	10	0.19±0.03	0.11±0.03	0.12±0.02	0.15±0.02	0.81±0.02
治疗组	10	0.61±0.02	0.48±0.03	0.32±0.01	0.25±0.03	0.38±0.03
F值	?	22.962	19.281	16.035	16.859	15.364
P值	?	0.001	0.001	0.005	0.007	0.010

组别	例数	Cdk-4	Cdk-6	Cyclin D1	p21
对照组	10	0.31±0.04	0.19±0.01	0.15±0.03	0.47±0.02
模型组	10	0.79±0.04	0.76±0.05	0.66±0.03	0.12±0.01
治疗组	10	0.39±0.03	0.24±0.03	0.19±0.01	0.41±0.03
F值	?	19.205	15.331	20.749	15.668
P值	?	0.001	0.005	0.001	0.003

组别	例数	Cdk-4	Cdk-6	Cyclin D1	p21
对照组	10	0.31±0.04	0.19±0.01	0.15±0.03	0.47±0.02
模型组	10	0.79±0.04	0.76±0.05	0.66±0.03	0.12±0.01
治疗组	10	0.39±0.03	0.24±0.03	0.19±0.01	0.41±0.03
F值	?	19.205	15.331	20.749	15.668
P值	?	0.001	0.005	0.001	0.003

组别	例数	OCT4	SOX2	c-Myc	TCF4
对照组	10	1.00±0.05	1.00±0.05	1.00±0.05	1.00±0.05
模型组	10	2.82±0.08	2.21±0.08	3.12±0.22	3.96±0.65
治疗组	10	1.34±0.02	1.36±0.02	1.15±0.01	1.11±0.01
F值	?	13.762	16.375	20.236	15.766
P值	?	0.010	0.016	0.014	0.011

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