Effect of RNAi Silencing of The DLL4 Gene for Inducing Apoptosis and Chemosensitivity to Docetaxel of Human Breast Cancer Cells_CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY

Authors：

WANG Qun ¹ , YU Minghua ² , WANG Geng ¹ ,  WANG Minghua ¹

1.Department of Endocrine and Vascular Surgery， Taihe Hospital， Hubei Medical University of Medicine， Shiyan 442000， Hubei Province， China;;
2.Institute of Basic Medical Sciences， Hubei Medical University of Medicine， Shiyan 442000， Hubei Province， China;

Corresponding author：

WANG Minghua, Email: wangmh1970@sina.com

Keywords：

RNA interference; Breast cancer; Delta-like ligand 4; Chemosensitivity

Video：

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Abstract Full text Figures/Tables Video References Cited by

Objective 　To explore effect of DLL4 gene in MCF-7 cells of human breast cancer which was inhibitted by short hair in RNA （shRNA） on inducing apoptosis and chemosensitivity to docetaxel.
Methods 　Specific shRNA was designed in accordance with DLL4 gene and transfected into MCF-7 cells of human breast cancer with liposomal （Lip-shRNA group）， MCF-7 cells transfected with only liposomal as Lip group， and control group without any treatment. Expressions of DLL4 protein in 3 groups were detected by immunohistochemical method， apoptosis and cell cycle distribution were examined by flow cytometry （FCM）. Proliferations and sensitivity of MCF-7 cells to docetaxel in 3 groups were determined by methylthiazoyl-tetrazolium bromide （MTT）.
Results 　The averages optical density value and rate of positive area of Lip-shRNA group were significantly lower than that of other 2 groups （P＜0.01）. The levels of A value at 24h， 48h， and 72h in Lip-shRNA group were significantly lower than that of other 2 groups （P＜0.01）. Rates of cell apoptosis at 24h， 48h， 72h and 96 h in Lip-shRNA group were significantly higher than that of other 2 goups （P＜0.05），and ratio of G2/M was higher too （P＜0.05）. IC50 of Lip-shRNA group to docetaxel was significantly lower than that of other 2 groups （P＜0.05）.
Conclusions 　The RNA interference technology can effectively block the expression of DLL4 gene. Inhibition of DLL4/Notch signaling pathway can lead to proliferation inhibition of cancer cell and a concomitant increase in cells undergoing apoptosis， and can enhance the cell sensitivity to docetaxel. DLL4 may be an important target for therapeutic approach of breast cancer.

Citation： WANG Qun,YU Minghua,WANG Geng,WANG Minghua. Effect of RNAi Silencing of The DLL4 Gene for Inducing Apoptosis and Chemosensitivity to Docetaxel of Human Breast Cancer Cells. CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY, 2013, 20(1): 54-59. doi: Copy

1.	Patel NS， Li JL， Generali D， et al. Up-regulation of delta-like 4 ligand in human tumor vasculature and the role of basal expression in endothelial cell function[J]. Cancer Res， 2005， 65(19)：8690-8697.
2.	Benedito R， Trindade A， Hirashima M， et al. Loss of notch signalling induced by Dll4 causes arterial calibre reduction by increasing endothelial cell response to angiogenic stimuli[J]. BMC Dev Biol， 2008， 8(16)：117.
3.	Li JL， Sainson RC， Shi W， et al. Delta-like 4 Notch ligand regulates tumor angiogenesis， improves tumor vascular function， and promotes tumor growth in vivo[J]. Cancer Res， 2007， 67(23)：11244-11253.
4.	Shin S， Sung BJ， Cho YS， et al. An anti-apoptotic protein humansurvivin is a direct inhibitor of caspase-3 and -7[J]. Biochemistry，2001， 40(4)：1117-1123.
5.	魏爱英，张庆柱，张捷. 多西他赛诱导乳腺癌Bcap37细胞凋亡作用的研究[J]. 中华肿瘤防治杂志， 2006， 13(18)：1384-1386.
6.	Siekmann AF， Lawson ND. Notch signalling limits angiogenic cell behaviour in developing zebrafish arteries[J]. Nature， 2007， 445(7129)：781-784.
7.	Scehnet JS， Jiang W， Kumar SR， et al. Inhibition of Dll4-mediated signaling induces proliferation of immature vessels and results in poor tissue perfusion[J]. Blood， 2007， 109(11)：4753-4760.
8.	Hellström M， Phng LK， Hofmann JJ， et al. Dll4 signalling through Notch1 regulates formation of tip cells during angiogenesis[J]. Nature， 2007， 445(7129)：776-780.
9.	Segarra M， Williams CK， Sierra Mde L， et al. Dll4 activation of Notch signaling reduces tumor vascularity and inhibits tumor growth[J]. Blood， 2008， 112(5)：1904-1911.
10.	Dufraine J， Funahashi Y， Kitajewski J. Notch signaling regulates tumor angiogenesis by diverse mechanisms[J]. Oncogene， 2008， 27(38)：5132-5137.
11.	Harrington LS， Sainson RC， Williams CK， et al. Regulation of multiple angiogenic pathways by Dll4 and Notch in humanumbilical vein endothelial cells[J]. Microvasc Res， 2008， 75(2)：144-154.
12.	Zang S， Ji CH，Qu X， et al. A study on Notch signaling in humanbreast cancer[J]. Neoplasma， 2007， 54(4)：304-310.
13.	Jubb AM， Soilleux EJ， Turley H， et al. Expression of vascular notch ligand delta-like 4 and inflammatory markers in breast cancer[J]. Am J Pathol， 2010， 176(4)：2019-2028.
14.	王群，余明华，王耕，等. DLL4和S100A4在不同分子亚型乳腺癌组织中的表达及意义[J]. 中国普外基础与临床杂志， 2012， 19(9)：46-50.
15.	王群，余明华，王耕，等. DLL4、COX-2和MMP-2在乳腺癌组织中的表达及临床意义[J]. 临床肿瘤学杂志， 2012， 17(9)：814-817.
16.	Indraccolo S， Minuzzo S， Masiero M， et al. Cross-talk between tumor and endothelial cells involving the Notch3-Dll4 interaction marks escape from tumor dormancy[J]. Cancer Res， 2009， 69(4)：1314-1323.
17.	Hoey T， Yen WC， Axelrod F， et al. DLL4 blockade inhibitstumor growth and reduces tumor-initiating cell frequency[J]. Cell Stem Cell， 2009， 5(2)：168-177.
18.	Kajiyama H， Shibata K， Terauchi M， et al. Chemoresistance to paclitaxel induces epithelial-mesenchymal transition and enhances metastatic potential for epithelial ovarian carcinoma cells[J]. Int J Oncol， 2007， 31(2)：277-283.
19.	Shah AN， Summy JM， Zhang J， et al. Development and characterization of gemcitabine-resistant pancreatic tumor cells[J]. Ann Surg Oncol， 2007， 14(12)：3629-3637.
20.	Wang Z， Azmi AS， Ahmad A， et al. TW-37， a small-molecule inhibitor of Bcl-2， inhibits cell growth and induces apoptosis in pancreatic cancer：involvement of Notch-1 signaling pathway[J]. Cancer Res， 2009， 69(7)：2757-2765.
21.	Min C， Eddy SF， Sherr DH， et al. NF-kappaB and epithelial to mesenchymal transition of cancer[J]. J Cell Biochem， 2008， 104(7)：733-744.
22.	Rye PD， Stigbrand T. Interfering with cancer：a brief outline of advances in RNA interference in oncology[J]. Tumour Biol， 2004， 25(5-6)：329-336.
23.	Hannon GJ. RNA interference[J]. Nature， 2002， 418(6894)：244-251.
24.	Brummelkamp TR， Bernards R， Agami R. A system for stable expression of short interfering RNAs in mammalian cells[J]. Science， 2002， 296(5567)：550-553.
25.	刘启志，王烈. siRNA表达载体对肝素酶基因的沉默作用[J]. 中国普外基础与临床杂志， 2010， 17(2)：142-146.

1. Patel NS， Li JL， Generali D， et al. Up-regulation of delta-like 4 ligand in human tumor vasculature and the role of basal expression in endothelial cell function[J]. Cancer Res， 2005， 65(19)：8690-8697.
2. Benedito R， Trindade A， Hirashima M， et al. Loss of notch signalling induced by Dll4 causes arterial calibre reduction by increasing endothelial cell response to angiogenic stimuli[J]. BMC Dev Biol， 2008， 8(16)：117.
3. Li JL， Sainson RC， Shi W， et al. Delta-like 4 Notch ligand regulates tumor angiogenesis， improves tumor vascular function， and promotes tumor growth in vivo[J]. Cancer Res， 2007， 67(23)：11244-11253.
4. Shin S， Sung BJ， Cho YS， et al. An anti-apoptotic protein humansurvivin is a direct inhibitor of caspase-3 and -7[J]. Biochemistry，2001， 40(4)：1117-1123.
5. 魏爱英，张庆柱，张捷. 多西他赛诱导乳腺癌Bcap37细胞凋亡作用的研究[J]. 中华肿瘤防治杂志， 2006， 13(18)：1384-1386.
6. Siekmann AF， Lawson ND. Notch signalling limits angiogenic cell behaviour in developing zebrafish arteries[J]. Nature， 2007， 445(7129)：781-784.
7. Scehnet JS， Jiang W， Kumar SR， et al. Inhibition of Dll4-mediated signaling induces proliferation of immature vessels and results in poor tissue perfusion[J]. Blood， 2007， 109(11)：4753-4760.
8. Hellström M， Phng LK， Hofmann JJ， et al. Dll4 signalling through Notch1 regulates formation of tip cells during angiogenesis[J]. Nature， 2007， 445(7129)：776-780.
9. Segarra M， Williams CK， Sierra Mde L， et al. Dll4 activation of Notch signaling reduces tumor vascularity and inhibits tumor growth[J]. Blood， 2008， 112(5)：1904-1911.
10. Dufraine J， Funahashi Y， Kitajewski J. Notch signaling regulates tumor angiogenesis by diverse mechanisms[J]. Oncogene， 2008， 27(38)：5132-5137.
11. Harrington LS， Sainson RC， Williams CK， et al. Regulation of multiple angiogenic pathways by Dll4 and Notch in humanumbilical vein endothelial cells[J]. Microvasc Res， 2008， 75(2)：144-154.
12. Zang S， Ji CH，Qu X， et al. A study on Notch signaling in humanbreast cancer[J]. Neoplasma， 2007， 54(4)：304-310.
13. Jubb AM， Soilleux EJ， Turley H， et al. Expression of vascular notch ligand delta-like 4 and inflammatory markers in breast cancer[J]. Am J Pathol， 2010， 176(4)：2019-2028.
14. 王群，余明华，王耕，等. DLL4和S100A4在不同分子亚型乳腺癌组织中的表达及意义[J]. 中国普外基础与临床杂志， 2012， 19(9)：46-50.
15. 王群，余明华，王耕，等. DLL4、COX-2和MMP-2在乳腺癌组织中的表达及临床意义[J]. 临床肿瘤学杂志， 2012， 17(9)：814-817.
16. Indraccolo S， Minuzzo S， Masiero M， et al. Cross-talk between tumor and endothelial cells involving the Notch3-Dll4 interaction marks escape from tumor dormancy[J]. Cancer Res， 2009， 69(4)：1314-1323.
17. Hoey T， Yen WC， Axelrod F， et al. DLL4 blockade inhibitstumor growth and reduces tumor-initiating cell frequency[J]. Cell Stem Cell， 2009， 5(2)：168-177.
18. Kajiyama H， Shibata K， Terauchi M， et al. Chemoresistance to paclitaxel induces epithelial-mesenchymal transition and enhances metastatic potential for epithelial ovarian carcinoma cells[J]. Int J Oncol， 2007， 31(2)：277-283.
19. Shah AN， Summy JM， Zhang J， et al. Development and characterization of gemcitabine-resistant pancreatic tumor cells[J]. Ann Surg Oncol， 2007， 14(12)：3629-3637.
20. Wang Z， Azmi AS， Ahmad A， et al. TW-37， a small-molecule inhibitor of Bcl-2， inhibits cell growth and induces apoptosis in pancreatic cancer：involvement of Notch-1 signaling pathway[J]. Cancer Res， 2009， 69(7)：2757-2765.
21. Min C， Eddy SF， Sherr DH， et al. NF-kappaB and epithelial to mesenchymal transition of cancer[J]. J Cell Biochem， 2008， 104(7)：733-744.
22. Rye PD， Stigbrand T. Interfering with cancer：a brief outline of advances in RNA interference in oncology[J]. Tumour Biol， 2004， 25(5-6)：329-336.
23. Hannon GJ. RNA interference[J]. Nature， 2002， 418(6894)：244-251.
24. Brummelkamp TR， Bernards R， Agami R. A system for stable expression of short interfering RNAs in mammalian cells[J]. Science， 2002， 296(5567)：550-553.
25. 刘启志，王烈. siRNA表达载体对肝素酶基因的沉默作用[J]. 中国普外基础与临床杂志， 2010， 17(2)：142-146.

CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY

Effect of RNAi Silencing of The DLL4 Gene for Inducing Apoptosis and Chemosensitivity to Docetaxel of Human Breast Cancer Cells

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