Effects of live attenuated measles vaccine Hu191 strain on epithelial mesenchymal transition,proliferation and migration of 4T1 breast cancer cells / 中国生物制品学杂志

@#Objective To investigate the effects of live attenuated measles vaccine Hu191 strain(MV-Hu191)on epithelial mesenchymal transition(EMT),proliferation and migration of 4T1 breast cancer cells.MethodsCCK-8 and clone formation assay were used to analyze the effect of MV-Hu191 on the proliferation of 4T1 cells;The effect of MV-Hu191 on 4T1cell migration was analyzed by cell scratch test;The expression of EMT pathway proteins(MMP-2,MMP-9,E-cadherin)in4T1 cells was detected by Western blot;4T1 tumor-bearing mouse model was established in female BALB/c mice. The model mice were divided into control group(PBS),MV-Hu191(1 × 106TCID50)group and paclitaxel group(15 mg/kg),with 10 mice in each group,and injected into tumor at the dosage of 100 μL every 2 d for 5 times. At 28 d after administration,the effects of MV-Hu191 on survival time,tumorigenicity and metastasis in vivo were observed;The pathological characteristics of lung tissue and tumor tissue were observed by HE staining under microscope;The expression of EMT pathway proteins(MMP-2,MMP-9 and E-cadherin)in tumor tissue was detected by immunohistochemical staining.Results The results of in vitro experiment showed that,compared with the control group,MV-Hu191 inhibited the proliferation and migration of 4T1 cells(F = 2. 811 and 13. 535,P = 0. 001 and 0. 002,respectively),down regulated the expression of MMP-2 and MMP-9(F = 45. 433 and 9. 744,P = 0. 011 and 0. 038,respectively),and up regulated the expression of Ecadherin(F = 7. 001,P = 0. 032);The results of in vivo experiment showed that MV-Hu191 significantly prolonged the survival time of tumor-bearing mice,and decreased the tumor quality(F = 8. 301,P = 0. 003)and the number of pulmonary nodules metastasis compared with the control group(F = 33. 792,P = 0. 000);MV-Hu191 treated tumor tissue gap was small,the cells were round,and the alveolar contour was clearly visible;The expression of MMP-2 and MMP-9 in MVHu191 treated tumor tissue decreased significantly(F = 6. 705 and 9. 047,P = 0. 028 and 0. 023,respectively),while the expression of E-cadherin increased significantly(F = 3. 468,P = 0. 039).ConclusionMV-Hu191 signi-ficantly inhibits the proliferation and migration of 4T1 breast cancer cells,antagonizes the tumorigenicity and lung meta-stasis of 4T1 tumorbearing mice,and prolongs the survival time of mice. The possible mechanism of MV-Hu191 against breast cancer is closely related to the regulation of EMT pathway protein expression.

Effect of Panax Notoginseng Saponins on Breast Cancer Cell Line 4T1 in Tumor-bearing Mice Through MEKK1/SEK1/JNK1/AP-1 Pathway / 中国实验方剂学杂志

Objective:To explore the potential mechanisms of Panax Notoginseng Saponins （PNS） on growth inhibition of breast cancer cell line 4T1 in tumor-bearing mice by investigating the mitogen-activated protein kinase kinase kinase 1 （MEKK1）/stress activated protein kinase （SAPK）/extracellular regulated protein kinases （Erk） Kinase （SEK1）/c-Jun N-terminal kinase 1 （JNK1）/activator protein-1 （AP-1） signaling pathways. Method:The 4T1 breast cancer mice model was established. Forty-eight mice with successful modeled and randomly divided into the low， medium and high-dose PNS groups （10， 20， 40 mg·kg-1） and the model control group （12 mice in each group）. The PNS groups received intraperitoneal injection with dosage of 10 mL·kg-1， while the controlled group was given the same dosage of saline. After administration with PNS for 28 days， tumor tissues were isolated， weighed， sliced and homogenized. Tumor cell apoptosis was detected by TdT mediated-dUTP nick end labeling （TUNEL） staining. The mRNA expressions of MEKK1， SEK1， JNK1 and AP-1 in tumor tissue were detected by Real-time polymerase chain reaction（Real-time PCR）. The protein expressions of MEKK1， SEK1， JNK1 and AP-1 in tumor tissue were detected by immunofluorescence staining and Western blot. Result:Compared with model group， the tumor weights of medium-dose and high-dose PNS groups were decreased significantly （P<0.05）. TUNEL staining showed that the number of apoptotic tumor cells increased with the rise of dosage of PNS （P<0.05）. The medium-dose and high-dose PNS groups showed a significant increase in the mRNA expressions of MEKK1， SEK1， JNK1 and AP-1 as well as the protein expressions of MEKK1， SEK1， JNK1 and AP-1 in tumor tissues （P<0.05）， with statistically significant differences （P<0.05）. Conclusion:PNS could inhibit the tumor growth of breast cancer cell line 4T1 in tumor-bearing mice， which may be related to the activation of MEKK1/SEK1/JNK1/AP-1 signaling pathways.

Total alkaloids of Coptidis Rhizoma combined with exercise inhibits tumor growth of orthotopically transplanted 4T1 breast cancer mice by blocking cell cycle G_1/S transformation / 中国中药杂志

Breast cancer is one of the leading causes for cancer-related death among women worldwide. Coptidis Rhizoma has antibacterial,anti-inflammatory,anti-tumor and other pharmacological activities,but whether exercise could synergistically promote the role of RC in the treatment of breast cancer has not been reported. In this experiment,the effects and mechanism of total alkaloids of Coptidis Rhizoma combined with exercise on the tumor growth of orthotopically transplanted 4 T1 breast cancer were systemically studied in mice. Balb/C mice transplanted with 4 T1 cells in situ were used as models. The total alkaloids of RC(145 mg·kg-1·d-1) alone or in combination with exercise(10 m·min-1,30 min/time,5 times/week) were given for 28 days,and then the changes in body weight and tumor volume,tumor weight,interleukin-1β(IL-1β),serum estradiol(E2) content,and expression levels of estrogen receptor α(ERα),cell cycle related proteins CDK4,CDK6,cyclin D1,CDK2,and cyclin E in tumor tissues. The results showed that total alkaloids of Coptidis Rhizoma could significantly inhibit the growth of 4 T1 breast cancer in mice(P< 0. 01),and exercise significantly promoted the anti-tumor activity of total alkaloids of Coptidis Rhizoma(P<0. 01),and reduced E2 and IL-1β levels in mice. Western blot and flow cytometry showed that the total alkaloids of Coptidis Rhizoma combined with exercise could down-regulate the protein expression levels of ERα,CDK4,CDK6,cyclin D1,CDK2 and cyclin E in cancer cells,block the transformation of G1/S in 4 T1 cell cycle,and inhibit DNA synthesis in breast cancer cells. The total alkaloids of Coptidis Rhizoma combined with exercise showed synergistic effect in inhibition of tumor growth in mice with orthotopically transplanted 4 T1 breast cancer.

Effects of polysaccharides from Ginkgo biloba on proliferation of 4T1 breast cancer cells and expression of GLUT family genes / 中国药理学通报

Aim To investigate the effects of polysac-charides from Ginkgo biloba on the proliferation, apop-tosis of mouse 4T1 breast cancer cells and the possible mechanism. Methods 4T1 cells in logarithmic growth phase were treated with polysaccharides from Ginkgo biloba of different concentrations. The effect of poly-saccharides from Ginkgo biloba on inhibition of prolif-eration and cytotoxicity of 4T1 cells was determined by MTT assay and trypan blue exclusion assay respective-ly. The apoptotic effect of polysaccharides from Ginkgo biloba on 4T1 cells was detected by DAPI staining. qRT-PCR experiments were carried out for the detec-tion of gene expressions of the glucose transporter fami-ly upon the treatment with the polysaccharides from Ginkgo biloba. Results Polysaccharides from either Ginkgo biloba leaf or Ginkgo biloba exocarp significant-ly inhibited the proliferation of 4T1 cells in a dose-and time-dependent manner. Moreover, with the increasing doses of polysaccharides, cell viability decreased, ac-companied by the increased cell cytotoxicity and apop-tosis. qRT-PCR results showed that polysaccharides from Ginkgo biloba significantly reduced glucose trans-porter 1 gene expression. Conclusions Polysaccha-rides from Ginkgo biloba can both inhibit 4T1 cell pro-liferation and induce cell apoptosis, and by regulating glucose transporter family gene expression, it interfered with cell energy metabolism, which infers that the effects of cell proliferation inhibition as well the apopto-sis induction might be due to the regulation of glucose transporter family gene expression.

Experimental Study on Anti-tumor Prescription Screening and Therapeutic Effects of Shengma Combined with Zao Jiao-ci / 中国中医药信息杂志

Objective To screen the anti-tumor prescription (Shengzao Fang) of Shengma (Rhizoma Cimicifugae) combined with Zao Jiao-ci (Spina Gleditsiae), and evaluate its anti-tumor effects. Methods The lung metastatic model of 4T1 breast cancer and the 4T1 subcutaneous transplanted model were established, both models of mice were randomly divided into model group, Shengma group, Zao Jiao-ci group, Shengzao Fang first group, Shengzao Fang second group and Shengzao Fang third group, 10 mice for each group. The proportion of Shengzao Fang was ascertained by their anti-tumor effects. MTT assay was used to investigate the effects of Shengzao Fang on 4T1 breast cancer. The concentration of type 1 collagen (Col-Ⅰ), platelet-activating factor (PAF), tissue inhibitor of metalloproteinase 1 (TIMP-1), tyrosine kinase receptor A (TrkA) and tyrosine kinase receptor B (TrkB) in serum from tumor-bearing mice was tested by enzyme-linked immunosorbent assay (ELISA). The effects of Shengzao Fang on matrix metalloproteinase (MMPs) was examined by Gelatin matrix method. Results Compared with the model group, the prescription of Shengma∶Zao Jiao-Ci=2∶1 significantly suppressed lung metastasis of 4T1 breast cancer, showing fewer lung nodes, lower lung metastasis rate and highest tumor inhibitory rate to 4T1 subcutaneous transplanted model. Although the concentration of serum Col-Ⅰ, PAF, TIMP-1, TrkA and TrkB was decreased in all treated group, the prescription of Shengma∶Zao Jiao-ci=2∶1 had the strongest activity, and its inhibitory effect on the expression of MMP-2 and MMP-9 was strongest, too. Conclusion The prescriptions of Shengma combined with Zao Jiao-ci had different antitumor activity, the strongest activity was exhibited when shengma∶Zao Jiao-ci was 2∶1, suggesting that 2∶1 was optimization for Shengma combined with Zao Jiao-ci.

Effect of resveratrol on the metastasis of 4T1 mouse breast cancer cells in vitro and in vivo

We investigated the effects of resveratrol on metastasis in in vitro and in vivo systems. 4T1 cells were cultured in the presence of various concentrations (0-30 micromol/L) of resveratrol. For experimental metastasis, BALB/c mice were injected intravenously with 4T1 cells in the tail vein, and were orally administered various concentrations (0, 100, or 200 mg/kg Body weight) of resveratrol for 21 days. After resveratrol treatment, cell adhesion, wound migration, invasion, and MMP-9 activity were significantly decreased in a dose-dependent manner in 4T1 cells (P < 0.05). The numbers of pulmonary nodules were significantly decreased in mice fed the resveratrol (P < 0.05). The plasma MMP-9 activity was decreased in response to treatment with resveratrol in mice (P < 0.05). We conclude that resveratrol inhibits cancer metastasis both in vitro and in vivo, and this inhibition is likely due to the decrease in MMP-9 activity caused by resveratrol.