Screening and Verification of RNA-seq-based PBCRS for Treating Key Genes of Breast Cancer
10.13422/j.cnki.syfjx.20190219
- VernacularTitle: 基于RNA-seq技术的活血化瘀法治疗乳腺癌关键基因的筛选和验证
- Author:
Li-nan ZHAO
1
;
Yi ZHAO
2
;
You-zhi SUN
2
Author Information
1. Chinese Medicine Hospital of Puyan, Puyang 457001, China
2. Jiangxi University of Traditional Chinese Medicine, Nanchang 330004, China
- Publication Type:Research Article
- Keywords:
breast cancer;
RNA-seq;
promoting blood circulation and removing stasis;
fructose-1,6-bisphosphatase1(FBP1);
purkinje cell protein 4(PCP4);
myoglobin(MB)
- From:
Chinese Journal of Experimental Traditional Medical Formulae
2019;25(4):101-108
- CountryChina
- Language:Chinese
-
Abstract:
Objective: To study the inhibitory effect of the medicine group of promoting blood circulation and removing stasis (PBCRS) on breast cancer induced by 7, 12-dimethylbenz(a)anthracene (DMBA) in rats, and screen out and verify key genes based on RNA Sequencing (RNA-seq) technology and Ingenuity Pathway Analysis (IPA). Method: Totally 96 Sprague-Dawley (SD) rats were randomly divided into blank group, DMBA model control group, tamoxifen (TAM) group (1.9 mg·kg-1·d-1), high-dose, middle-dose and low-dose PBCRS groups (12.96, 6.48, 3.24 g·kg-1·d-1). One week after drug intervention, except for the blank group, the DMBA was used to induce the rat model of breast cancer (with an interval of a week, irrigation for two times at the dose of 100 mg·kg-1). After 10 weeks, the changes in tumor weight and tumor volume were observed. The total RNA was extracted by total RNA extraction kit, and three RNA samples were collected from the DMBA model control group and the middle-dose PBCRS group for genetic testing. Based on RNA-seq, key differential genes were screened out and verified by Real-time PCR. Result: Comparing with the DMBA model control group, the tumor volume and tumor weight in middle-dose PBCRS group were decreased significantly (P<0.05). Although the tumor weight and tumor volume in high and low-dose PBCRS groups were decreased, there was no statistically significant difference. Based on RNA-seq technology and IPA analysis software, fructose-1, 6-bisphosphatase1 (FBP1), purkinje cell protein 4 (PCP4), myoglobin (MB) key genes were screened out. Compared with the DMBA model control group, the mRNA expressions of FBP1 in the high and middle-dose PBCRS groups were significantly increased (P<0.05). Although the mRNA expressions of PCP4 in the high, middle and low-dose PBCRS groups were increased, there was no statistical significance. The mRNA expression of MB in the low-dose group was decreased, but there was no statistical significance. Conclusion: PBCRS may inhibit the occurrence of breast cancer by interfering with the expression of FBP1 in breast cancer tissue.
