RNA-Seq Analysis of Cordycepin Against Triple-negative Breast Cancer

doi:10.13560/j.cnki.biotech.bull.1985.2021-0444

Abstract

Abstract:

Previous studies have shown that cordycepin,the active ingredient of Cordyceps militaris,can effectively inhibit the growth and metastasis of triple-negative breast cancer xenografts tumor,however its mechanism is not clear yet,thus the purpose of this paper is to predict its mechanism. Taking samples of triple-negative breast cancer xenografts tumor,the molecular mechanism of cordycepin against breast cancer xenografts tumor was analyzed through transcriptome sequencing technology. The results showed that 584 differentially expressed genes were screened using P<0.05,log₂|FC|>1 as the standard,including 69 up-regulated genes and 515 down-regulated genes. KEGG analysis demonstrated that cordycepin had molecular regulation on breast cancer xenografts tumors including Hippo signaling pathway,Hedgehog signaling pathway,ECM-receptor interaction,TGF-beta signaling pathway and other cancer-related signaling pathways. GO analysis revealed that these differential genes were enriched in 16 biological processes,such as biological adhesion,growth,metabolic process,and locomotion,etc. It is suggested that cordycepin had significant regulatory effect on the growth and metastasis of breast cancer xenografts tumors. Further protein-protein interaction analysis of these genes in the growth and metastasis process indicated that Hedgehog signaling pathway and Hippo signaling pathway were important potential ones for cordycepin against triple-negative breast cancer. The research results suggest that the Hippo-Yap/TAZ signaling pathway and the Hedgehog-Gli signaling pathway are potential ones,laying the foundation for exploring the molecular mechanism of cordycepin in the treatment of triple-negative breast cancer.

Key words: cordycepin, RNA-Seq, triple negative breast cancer, Hedgehog, Hippo

HUANG Hai-chen, WU Wen-ya, QI Meng, XUE Fan-zheng, WU Xiao-ping, ZHANG Jun-li, FU Jun-sheng. RNA-Seq Analysis of Cordycepin Against Triple-negative Breast Cancer[J]. Biotechnology Bulletin, 2021, 37(11): 72-80.

Figures/Tables 15

References 22

[1]	袁晓玲, 尤婧蓉, 沈莉莉, 等. 乳腺癌化学治疗患者更年期症状相关生活质量及影响因素研究[J]. 上海交通大学学报:医学版, 2018, 38(12): 1457-1462.
	Yuan XL, You JR, Shen LL, et al. Study on the predictors of the menopausal symptoms related quality of life in patients with breast cancer undergoing chemotherapy[J]. J Shanghai Jiao Tong Univ:Med Sci, 2018, 38(12): 1457-1462.
[2]	赵易, 赵庆丽, 马骥. 二甲双胍联合多西他赛对乳腺癌细胞MDA-MB-231增殖和凋亡的影响[J]. 现代肿瘤医学, 2018, 26(1): 10-13.
	Zhao Y, Zhao QL, Ma J. Proliferation and apoptosis effect of combining metformin and docetaxel on breast cancer cells[J]. J Mod Oncol, 2018, 26(1): 10-13.
[3]	倪晨, 李婷, 吴振华, 等. 三阴性乳腺癌化疗进展[J]. 中国癌症杂志, 2014, 24(4): 316-320.
	Ni C, Li T, Wu ZH, et al. Progress on chemotherapy for triple negative breast cancer[J]. China Oncol, 2014, 24(4): 316-320.
[4]	Auvinen P, Tammi R, Tammi M, et al. Expression of CD44s, CD44v3 and CD44v6 in benign and malignant breast lesions:correlation and colocalization with hyaluronan[J]. Histopathology, 2005, 47(4): 420-428. pmid: 16178897
[5]	Liu C, Qi M, Li L, et al. Natural cordycepin induces apoptosis and suppresses metastasis in breast cancer cells by inhibiting the Hedgehog pathway[J]. Food Funct, 2020, 11(3): 2107-2116. doi: 10.1039/C9FO02879J URL
[6]	Habib JG, O’Shaughnessy JA. The hedgehog pathway in triple-negative breast cancer[J]. Cancer Med, 2016, 5(10): 2989-3006. doi: 10.1002/cam4.2016.5.issue-10 URL
[7]	Pandolfi S, Stecca B. Cooperative integration between HEDGEHOG-GLI signalling and other oncogenic pathways:implications for cancer therapy[J]. Expert Rev Mol Med, 2015, 17: e5. doi: 10.1017/erm.2015.3 URL
[8]	Briscoe J, Thérond PP. The mechanisms of Hedgehog signalling and its roles in development and disease[J]. Nat Rev Mol Cell Biol, 2013, 14(7): 416-429. doi: 10.1038/nrm3598 URL
[9]	Tang JY, Mackay-Wiggan JM, Aszterbaum M, et al. Inhibiting the hedgehog pathway in patients with the basal-cell nevus syndrome[J]. N Engl J Med, 2012, 366(23): 2180-2188. doi: 10.1056/NEJMoa1113538 URL
[10]	Kayed H, Kleeff J, Osman T, et al. Hedgehog signaling in the normal and diseased pancreas[J]. Pancreas, 2006, 32(2): 119-129. doi: 10.1097/01.mpa.0000202937.55460.0c URL
[11]	Park JH, Shin JE, Park HW. The role of hippo pathway in cancer stem cell biology[J]. Mol Cells, 2018, 41(2): 83-92.
[12]	Cordenonsi M, Zanconato F, Azzolin L, et al. The hippo transducer TAZ confers cancer stem cell-related traits on breast cancer cells[J]. Cell, 2011, 147(4): 759-772. doi: 10.1016/j.cell.2011.09.048 URL
[13]	Maugeri-Saccà M, De Maria R. Hippo pathway and breast cancer stem cells[J]. Crit Rev Oncol Hematol, 2016, 99: 115-122. doi: 10.1016/j.critrevonc.2015.12.004 pmid: 26725175
[14]	Moroishi T, Hayashi T, Pan WW, et al. The hippo pathway kinases LATS1/2 suppress cancer immunity[J]. Cell, 2016, 167(6): 1525-1539.e17. doi: S0092-8674(16)31526-4 pmid: 27912060
[15]	Britschgi A, Duss S, Kim S, et al. The Hippo kinases LATS1 and 2 control human breast cell fate via crosstalk with ERα[J]. Nature, 2017, 541(7638): 541-545. doi: 10.1038/nature20829 URL
[16]	Ercolani C, Di Benedetto A, Terrenato I, et al. Expression of phosphorylated Hippo pathway kinases(MST1/2 and LATS1/2)in HER2-positive and triple-negative breast cancer patients treated with neoadjuvant therapy[J]. Cancer Biol Ther, 2017, 18(5): 339-346. doi: 10.1080/15384047.2017.1312230 URL
[17]	Heidary Arash E, Shiban A, Song S, et al. MARK4 inhibits Hippo signaling to promote proliferation and migration of breast cancer cells[J]. EMBO Rep, 2017, 18(3): 420-436. doi: 10.15252/embr.201642455 pmid: 28183853
[18]	Yao CB, Zhou X, Chen CS, et al. The regulatory mechanisms and functional roles of the Hippo signaling pathway in breast cancer[J]. Yi Chuan, 2017, 39(7): 617-629.
[19]	Sulaiman A, McGarry S, Li L, et al. Dual inhibition of Wnt and Yes-associated protein signaling retards the growth of triple-negative breast cancer in both mesenchymal and epithelial states[J]. Mol Oncol, 2018, 12(4): 423-440. doi: 10.1002/mol2.2018.12.issue-4 URL
[20]	Andrade D, Mehta M, Griffith J, et al. YAP1 inhibition radiosensitizes triple negative breast cancer cells by targeting the DNA damage response and cell survival pathways[J]. Oncotarget, 2017, 8(58): 98495-98508. doi: 10.18632/oncotarget.21913 pmid: 29228705
[21]	Cotton JL, Li Q, Ma L, et al. YAP/TAZ and hedgehog coordinate growth and patterning in gastrointestinal mesenchyme[J]. Dev Cell, 2017, 43(1): 35-47.e4. doi: 10.1016/j.devcel.2017.08.019 URL
[22]	Watanabe Y, Miyasaka KY, Kubo A, et al. Notch and Hippo signaling converge on Strawberry Notch 1(Sbno1)to synergistically activate Cdx2 during specification of the trophectoderm[J]. Sci Rep, 2017, 7(1): 1-17. doi: 10.1038/s41598-016-0028-x URL

基因Gene	引物序列Primer sequence（5'-3'）
PDK4	F:5'-GAGGATTACTGACCGCCTCTTTAG-3' R:5'-TTCCGGGAATTGTCCATCAC-3'
CD24	F:5'-CCCACGCAGATTTATTCCAG-3' R:5'-GACTTCCAGACGCCATTTG-3'
MMP7	F:5'-AGCCAAACTCAAGGAGATGC-3' R:5'-GCCAATCATGATGTCAGCAG-3'
SHH	F:5'-AGGGCACCATTCTCATCAAC-3' R:5'-GGAGCGGTTAGGGCTACTCT-3'
GLI2	F:5'-GCCCTT CCTGAAAAGAAGAC-3' R:5'-CATTGGAGA AACAGGATTGG-3'
GADPH	F:5'-GAAGGACTCATGACCACAG-3' R:5'-CTTCACCACCTTCTTGATG-3'

基因Gene	引物序列Primer sequence（5'-3'）
PDK4	F:5'-GAGGATTACTGACCGCCTCTTTAG-3' R:5'-TTCCGGGAATTGTCCATCAC-3'
CD24	F:5'-CCCACGCAGATTTATTCCAG-3' R:5'-GACTTCCAGACGCCATTTG-3'
MMP7	F:5'-AGCCAAACTCAAGGAGATGC-3' R:5'-GCCAATCATGATGTCAGCAG-3'
SHH	F:5'-AGGGCACCATTCTCATCAAC-3' R:5'-GGAGCGGTTAGGGCTACTCT-3'
GLI2	F:5'-GCCCTT CCTGAAAAGAAGAC-3' R:5'-CATTGGAGA AACAGGATTGG-3'
GADPH	F:5'-GAAGGACTCATGACCACAG-3' R:5'-CTTCACCACCTTCTTGATG-3'

样品 Sample	浓度 Concentration/（ng·μL^-1）	体积 Volume /μL	总量 Total /μg	RIN值 RIN value	文库类型 Library type	结论 Conclusion
C0R1	295	35	10.33	6.8	RNAseq	A
C0R2	528	35	18.46	7.9	RNAseq	A
C0R3	761	35	26.67	9.3	RNAseq	A
C0N1	652	76	49.55	8.3	RNAseq	A
C0N2	695	76	52.78	9.3	RNAseq	A
C0N3	602	74	44.51	9.3	RNAseq	A

样品 Sample	浓度 Concentration/（ng·μL^-1）	体积 Volume /μL	总量 Total /μg	RIN值 RIN value	文库类型 Library type	结论 Conclusion
C0R1	295	35	10.33	6.8	RNAseq	A
C0R2	528	35	18.46	7.9	RNAseq	A
C0R3	761	35	26.67	9.3	RNAseq	A
C0N1	652	76	49.55	8.3	RNAseq	A
C0N2	695	76	52.78	9.3	RNAseq	A
C0N3	602	74	44.51	9.3	RNAseq	A

样品名 Sample	总reads数量 Number of total reads	比对上rRNA的 reads 数及占总数的比例 Mapped reads（ratio in total）	未比对上rRNA的 reads 数及占总数的比例 Unmapped reads（ratio in total）
CON1	53195320	410412（0.77%）	52784908（99.23%）
CON2	69510386	493598（0.71%）	69016788（99.29%）
CON3	65958874	550350（0.83%）	65408524（99.17%）
COR1	90497008	812122（0.90%）	89684886（99.10%）
COR2	78992736	387314（0.49%）	78605422（99.51%）
COR3	56738684	310270（0.55%）	56428414（99.45%）