新型VEGF单克隆抗体与全反式维甲酸联合应用对乳腺癌MCF-7细胞增殖的抑制作用

doi:10.13481/j.1671-587X.20210213

Abstract

Abstract: Objective

To explore the effects of new vascular endothelial growth factor （VEGF） monoclonal antibody， all-trans retinoic acid （ATRA）， new VEGF monoclonal antibody combined with ATRA on the breast cancer MCF-7 cells， and to provide a new clinical approach for the treatment of breast cancer.

Methods

The breast cancer MCF-7 cells were divided into control group （without any treatment）， VEGF antibody group （20 mg·L^－1VEGF antibody）， ATRA group （10 μmol·L^－1 ATRA） and VEGF antibody + ATRA group （20 mg·L^－1 VEGF antibody+10 μmol·L^－1 ATRA）， each with 5 replicates. After the MCF-7 cells were treated with different drugs for 48 h，the VEGF antibody concentration was determined by ELISA， the proliferation rate of MCF-7 cells was determined by MTT assay， the apoptotic rate of MCF-7 cells was determined by flow cytometry， and the expression levels of B cell lymphoma-2（Bcl-2），Bcl-2 associated X protein（ Bax） and nuclear factor-κB（NF-κB） proteins in the MCF-7 cells were determined by Western blotting method.

Results

The ELISA results showed that the new human recombinant VEGF monoclonal antibody had good quality compared with positive antibody control and its concentration was 200 μg·L^-1.The MTT results showed that the prolifenation rate of MCF-7 cells in VEGF antibody + ATRA group was decreased obviously compared with control group （P<0.01）.The flow cytometry results showed that compared with control group，VEGF antibody group and ATRA group， the apoptotic rate of MCF-7 cells in VEGF antibody + ATRA group was significantly increased （P<0.01）；compared with control group，the apoptotic rates of MCF-7 cells in VEGF antibody group and ATRA group were significantly increased（P<0.01）；compared with VEGF antibody group，the apototic rate of MCF-7 cells in ATRA group was significantly increased（P<0.01）. The results of Western blotting method showed that compared with control group， VEGF antibody group and ATRA group， the expression level of Bcl-2 protein in VEGF antibody+ATRA group was significantly decreased （P<0.05）；compared with control group and VEGF antibody group， the expression level of NF-κB protein in MCF-7 cells in ATRA group was decreased （P<0.05 or P<0.01）， and the expression level of Bax protein was increased （P<0.01）.

Conclusion

The new VEGF monoclonal antibody and ATRA can inhibit the cell proliferation and promote apoptosis of breast cancer MCF-7 cells， and the effect of their combination is significantly better than each drug used alone.

Key words: breast neoplasms, vascular endothelial growth factor, monoclonal antibody, all-trans retinoic acid, apoptosis, proliferation

CLC Number:

R737.9

Haoxuan TANG,Nanfeng MENG,Meiling DU,Wei WANG,Tao HE. Inhibitory effect of new VEGF monoclonal antibody combined with all-trans retinoic acid on proliferation of breast cancer MCF-7 cells[J].Journal of Jilin University(Medicine Edition), 2021, 47(2): 344-351.

Figures/Tables 8

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References 29

1	HU J， LI G， ZHANG P， et al.A CD44v+ subpopulation of breast cancer stem-like cells with enhanced lung metastasis capacity［J］. Cell Death Dis， 2017， 8（3）：e2679.
2	CUSTÓDIO-SANTOS T，VIDEIRA M，BRITO M A.Brain metastasization of breast cancer［J］. Biochim Biophys Acta Rev Cancer， 2017， 1868（1）： 132-147.
3	BRAY F， FERLAY J， SOERJOMATARAM I， et al. Global cancer statistics 2018： GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries［J］. CA Cancer J Clin， 2018， 68（6）： 394-424.
4	CHEN W， ZHENG R， BAADE P D， et al. Cancer statistics in China， 2015［J］. CA Cancer J Clin， 2016， 66（2）：115-132.
5	BOLIS M， GARATTINI E， PARONI G， et al. Network-guided modeling allows tumor-type independent prediction of sensitivity to all-trans-retinoic acid［J］. Ann Oncol， 2017， 28（3） 611-621.
6	FOX S B，GENERALI D G，HARRIS A L.Breast tumour angiogenesis［J］. Breast Cancer Res，2007， 9（6）：216.
7	MASI A， LEBOFFE L， DE M E，et al. Retinoic acid receptors： from molecular mechanisms to cancer therapy［J］. Mol Aspects Med，2015， 41：1-115.
8	JUSSILA L ， VALTOLA R， PARTANEN T A. et al. Lymphatic endothelium and Kaposi’s sarcoma spindle cells detected by antibodies against the vascular endothelial growth factor receptor-3［J］. Cancer Res，1998，58：1599-1604.
9	SKOBE M，HAWIGHORST T，JACKSON D G，et al.Induction of tumor lymphangiogenesis by VEGF-C promotes breast cancer metastasis［J］.Nat Med， 2001，7（2）：192-198.
10	KIM， K J， LI B， WINER J， et al. Inhibition of vascular endothelial growth factor induced angiogenesis suppresses tumour growth in vivo［J］.Nature，1993， 362（6423）： 841-844.
11	XU Q H，SUN T，TIAN H，et al．Ultrasound-mediated vascular endothelial growth factor C（VEGF-C）gene microbubble transfection inhibits growth of MCF-7 breast cancer cells［J］.Oncol Res，2013，20（7）：297-301．
12	PAPADOPOULOS N， MARTIN J， RUAN Q， et al. Binding and neutralization of vascular endothelial growth factor （VEGF） and related ligands by VEGF Trap，ranibizumab and bevacizumab［J］. Angiogenesis， 2012， 15（2）：171-185.
13	VAZIRI S， KIM J， GANAPATHI M K，et al. Vascular endothelial growth factor polymorphisms：role in response and toxicity of tyrosine kinase inhibitors［J］. Curr Oncol Rep， 2010， 12（2）：102-108.
14	VISHAL R，SANJAYKUMAR H，SHENHONG W.et al.Treatment-related mortality with bevacizumab in cancer patients［J］. JAMA， 2011，305（5）：487-494.
15	董雪，徐雨昕，郭晶，等.重组人源抗血管内皮生长因子单克隆抗体的制备及鉴定［J］.吉林大学学报（理学版），2019，57（6）：1538-1543.
16	薛文潮，李光，张妮娜，等.全反式维甲酸联合辛二酰苯胺异羟肟酸诱导MCF-7细胞凋亡［J］.现代生物医学进展，2016，16（20）：3845-3848.
17	LISA R， LINDERHOLM B， NIELSEN N H， et al. Tumor specific VEGF-A and VEGFR2/KDR protein are co-expressed in breast cancer［J］. Breast Cancer Res Treat， 2003， 82（3）：147-154.
18	CHEN J Y，LAI Y S，CHU P Y，et al. Cancer-derived VEGF-C increases chemokine production in lymphatic endothelial cells to promote CXCR2-dependent cancer invasion and MDSC recruitment［J］. Cancers，2019，11（8）：1120.
19	NAGY J A， VASILE E， FENG D， et al. Vascular permeability factor/vascular endothelial growth factor induces lymphangiogenesis as well as angiogenesis［J］. J Exp Med， 2002， 196（11）：1497-1506.
20	YAMADA K， UCHIYAMA A， UEHARA A， et al. MFG-E8 drives melanoma growth by stimulating mesenchymal stromal cell-induced angiogenesis and M2 polarization of tumor-associated macrophages［J］. Cancer Res， 2016 76（14）：4283-4292.
21	ZHAO Z，LI Y K，SHUKLA R，et al. Development of a biocompatible copolymer nanocomplex to deliver VEGF siRNA for triple negative breast cancer［J］. Theranostics， 2019， 9（15）：4508-4524.
22	HUANG S Y，CHEN Y，LIANG Z M，et al. Targeting pin1 by all-trans retinoic acid （ATRA） overcomes tamoxifen resistance in breast cancer via multifactorial mechanisms.［J］. Front Cell Dev Biol， 2019， 7：322.
23	TERAO M，GORACCI L，CELESTINI V，et al. Role of mitochondria and cardiolipins in growth inhibition of breast cancer cells by retinoic acid.［J］. J Exp Clin Cancer Res， 2019， 38：496.
24	HUANG T Y， PENG S F， HUANG Y P，et al. Combinational treatment of all-trans retinoic acid （ATRA） and bisdemethoxycurcumin （BDMC）-induced apoptosis in liver cancer Hep3B cells［J］. J Food Biochem， 2020，44（2）：e13122.
25	李四强，侯忠赤，宫超，等. 全反式维甲酸对HL-60细胞NF-κB、CyclinD1表达的影响［J］. 中国现代医学杂志， 2006， 16（2）：180-182.
26	朱迎萍，吴燕平. 自拟宫颈方对宫颈炎模型大鼠宫颈组织VEGF-A、NF-KB表达的影响［J］. 浙江中西医结合杂志， 2017，27（9）：22-25.
27	曹文荣，闫文修，王雪利. 非小细胞肺癌组织中VEGF和Bax的表达及意义［J］. 重庆医学，2016（3）：307-309.
28	史爱英.VEGF表达下调抑制人宫颈癌细胞增殖转移相关机制研究［J］.中国实验诊断学，2019，23（10）：1816-1818.
29	GE Y L， ZHANG X， ZHANG J Y， et al. The mechanisms on apoptosis by inhibiting VEGF expression in human breast cancer cells［J］. Int Immunopharmacol， 2009， 9（4）：389-395.

Group	A value	Protein concentrasion[ρ_B/（μg·L^－1）]
VEGF-antibody	1.879±0.045	200
Positive antibody standard	1.119±0.060	125

Group	Cytotoxicity
Control	0.30±0.013
1 μmol·L^－1 ATRA+15 mg·L^－1 VEGF-antibody	0.32±0.011
5 μmol·L^－1 ATRA+15 mg·L^－1 VEGF-antibody	0.32±0.005
1 μmol·L^－1 ATRA+20 mg·L^－1 VEGF-antibody	0.34±0.011^*
5 μmol·L^－1 ATRA+20 mg·L^－1 VEGF-antibody	0.36±0.014^*
10 μmol·L^－1 ATRA+10 mg·L^－1 VEGF-antibody	0.36±0.020^*
10 μmol·L^－1 ATRA+15 mg·L^－1 VEGF-antibody	0.38±0.020^**
15 μmol·L^－1 ATRA+10 mg·L^－1 VEGF-antibody	0.43±0.018^**
10 μmol·L^－1 ATRA+20 mg·L^－1 VEGF-antibody	0.51±0.019^**

Group	Proliferation rate
Control	48.02±2.95
VEGF-antibody	37.26±1.11^*
ATRA	34.22±0.75^*
ATRA+VEGF-antibody	31.12±0.36^*△#

Group	Apoptotic rate
Control	6.25±1.28
VEGF-antibody	14.60±0.39^*
ATRA	19.75±1.82^*
ATRA+VEGF-antibody	31.35±1.73^*△#

Group	Bcl-2 protein	Bax protein	NF-κB protein
Control	0.139 4±0.007 6	0.191 9±0.016 4	0.174 9±0.017 9
VEGF-antibody	0.122 2±0.007 6^**	0.379 4±0.052 6^**	0.172 9±0.014 5
ATRA	0.096 5±0.005 8^**	0.646 0±0.065 5^**△△	0.117 6±0.004 9^*△△
ATRA+VEGF-antibody	0.079 0±0.006 0^**△△#	0.844 2±0.072 7^**△△	0.129 8±0.005 9^*△
P	<0.01	<0.01	<0.05

Inhibitory effect of new VEGF monoclonal antibody combined with all-trans retinoic acid on proliferation of breast cancer MCF-7 cells

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