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Objective:To study the efficacy and mechanism of Shugan Jianpi Jiedu prescription （SJJ） in the treatment of triple-negative breast cancer through <italic>in vitro</italic> cell experiments. Method:The following groups were set up in this study： a normal serum group，a pirarubicin group，and low-，medium-， and high-dose SJJ-medicated serum groups. Twenty SD rats were randomly divided into four groups and administered with SJJ solution （16.8，8.2，4.05 g·kg<sup>-1</sup>） and normal saline （equal volume） according to the body surface area to prepare serum. MDA-MB-231 cells were treated separately. The proliferation， migration and invasion of MDA-MB-231 cells were detected by the cell counting kit-8（CCK-8），wound healing assay and transwell cell invasion assay. The phosphoinositide 3-kinase （PI3K），protein kinase B （Akt）， and mechanistic target of rapamycin （mTOR） protein expression levels in MDA-MB-231 cells were tested by the Western blot. Result:The cell proliferation in the three different doses of medicated serum groups and the pirarubicin positive control group was significantly inhibited as compared with that in the normal serum group（<italic>P</italic><0.01），and there was no statistical difference for this between the medium/high dose medicated serum group and the pirarubicin positive control group.The wound healing in the SJJ-medicated serum groups and the pirarubicin group was slowed down as compared with that in the normal serum group （<italic>P</italic><0.01），and the effect in the SJJ-medicated serum groups was weaker than that in the pirarubicin group （<italic>P</italic><0.05，<italic>P</italic><0.01）. The number of cells invading the lower transwell chamber was decreased as compared with that in the normal serum group （<italic>P</italic><0.01），and there was no statistical difference between the medium-/high-dose SJJ-medicated serum groups and the pirarubicin group. Western blot results showed that 48 h after treatment，the PI3K，Akt， and mTOR expression levels in the cells of SJJ-medicated serum groups and the pirarubicin group were lower than those of the normal serum group（<italic>P</italic><0.01）. Conclusion:The SJJ-medicated serum could inhibit the proliferation， migration and invasion of MDA-MB-231 cells presumedly by down-regulating the protein expression levels in the PI3K/Akt/mTOR signaling pathway.
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@#[Abstract] Objective: To investigate the effect of lncRNA SNHG15 targeting miR-153 on cell viability and apoptosis of breast cancer cells and its apoptotic mechanism. Methods:The expression of SNHG15 in breast cancer cell lines(MDA-MB-231, BT-549 and MCF-7) were detected by Real-time fluorescent quantitative PCR (qPCR). MDA-MB-231 cells were divided into control (Ctrl) group, si-NC group, si-SNHG15 group, si-SNHG15+anti-NC group and si-SNHG15+anti-miR-153 group. Cell viability and apoptosis rate were detected by MTT and Flow cytometry, respectively. The targeting relationship between SNHG15 and miR-153 was verified by Dual luciferase report gene system. Mitochondrial membrane potential fluorescent probe (JC-1) staining method was used to detect cell mitochondrial membrane potential. The expressions of mitochondrial apoptosis-related proteins (Bcl-2, Bax, caspase3, cleaved caspase3 [c-caspase3] and Cyt-C）were detected by Western blotting. Results: The expression of SNHG15 in breast cancer cells was significantly higher than that in human normal mammary epithelial MCF10A cells (P<0.01). There was a targeting relationship between SNHG15 and miR-153. Compared with the control group, the cell viability and mitochondrial membrane potential of MDA-MB-231 cells in si-SNHG15 group were decreased, while apoptosis rate was increased (all P<0.01); the expressions of Bcl-2 and caspase3 were decreased while expressions of Bax, c-caspase3 and Cyt-C were increased (all P<0.01). However, co-transfection of si-SNHG15 and anti-miR-153 significantly attenuated the effects of si-SNHG15 on cell viability, apoptosis, mitochondrial membrane potential and expressions of Bcl-2, Bax, caspase3, c-caspase3 and Cyt-C (all P<0.01). Conclusion: lncRNA SNHG15 can target miR-153 to induce apoptosis of MDA-MB-231 cells, and the mechanism may be related to the regulation of apoptosis of mitochondrial pathway.
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@#Objective: To investigate the expression of tetraspanins-29 (Tspan29) in breast cancer tissues and cell lines and to explore the effect of Tspan29 knockdown on proliferation, invasion, migration and epithelial-mesenchymal transition (EMT) of breast cancer MCF-7 and MDA-MB-231 cells. Methods:Atotal of20pairsofbreast cancer tissues and corresponding para-cancerous tissues resected in Minhang Branch of Cancer HospitalAffiliated to Fudan University from June 2017 to February 2018 were collected for this study; in addition, breast cancer celllinesMCF-7,MDA-MB-231andhumanbreastepithelialMDA-kb2cellswerealsocollected.ThemRNAand protein expressions of Tspan29 in above mentioned tissues and cell lines were detected by Real-time quantitative (qPCR) and Western blotting. The expression of Tspan29 in MCF-7 and MDA-MB-231 cells was interfered by siRNA. qPCR was used to detect the mRNA and protein expressions of Tspan29. PCR microarray was used to examine the expressions of EMT-related genes in MCF-7 cells. CCK-8 assayandTranswellwereusedtodetectcellproliferation, migration and invasion of MCF-7 and MDA-MB-231 cells. Results: The mRNA and protein expressions of Tspan29 in breast cancer tissues were significantly higher than that in para-cancerous tissues (all P<0.01); and the mRNA and protein expressions of Tspan29 in MCF-7 and MDA-MB-231 cells were significantly higher than that in MDA-kb2 cells (P<0.01). After being interfered with siTspan29, the mRNA and protein expressions of Tspan29 were significantly down-regulated in MCF-7 cells (all P<0.05); the proliferation, invasion and migration of MCF-7 and MDA-MB-231 cells were significantly inhibited (all P<0.05); and among the EMT-related genes, two were significantly up-regulated while 7 were down-regulated. Conclusion: Tspan29 is significantly up-regulated in breast cancer tissues and cell lines, and knockdown of Tspan29 significantly inhibits the proliferation, invasion and migration of breast cancer cells. ··
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@#Objective: To investigate the effects of miR-200c on the proliferation, apoptosis and migration of triple negative breast cancer cell (TNBC) MDA-MB-231 and its metabolism-related molecular mechanism. Methods: miR-200c-231 (MDA-MB-231 overexpressing miR-200c) cells, miR-NC-231 (MDA-MB-231 transfected with miRNA-negative control) and the corresponding transplanted tumor models in nude mice were used as the study subjects. qPCR was used to detect the content of miR-200c and other related genes in cells and transplanted tumor tissues. The number of Ki67 positive cells in tumor tissue was analyzed by immunohistochemistry. The migration and apoptosis of cells were examined by Transwell chamber method and Flow cytometry, respectively. The expressions of proteins associated with proliferation, migration, and metabolism related signaling pathways in cells and tissues were confirmed by Western blotting. The changes of oxygen consumption rate (OCR), extracellular acidification rate (ECAR) and metabolic phenotype were detected by Seahorse energy metabolism detector. UPLC/LTQ-Orbitrap-MStechniquewasusedtoprofilethedifference of metabolites in cells. Results: The content of miR-200c in miR-200c-231cellswassignificantlyhigherthanthatinmiR-NC-231cells.Themass ofmiR-200c-231transplantedtumornotablydecreased,andthenumberofKi67positivecellsintumortissuesalsodecreasedsignificantly. The migration ability of miR-200c-231 cells decreased and the apoptosis rate increased (all P<0.01), accompanied with declined expressionsofZEB1/2,Vimentin,cyclinD1andincreasedexpressionofcleavedPARP(P<0.05orP<0.01),aswellasdecreasedphosphorylation leverofSTAT1/3andNF-κBbutincresedphosphorylationleverofCAMP(allP<0.05).OverexpressionofmiR-200cinMDA-MB-231cells increasedOCRandthecontentof10antitumor metabolites, but decreased ECAR and tryptophan 2,3-plus dioxidase (TDO2) expression (P<0.05 or P<0.01). Conclusion: miR-200c targeting TDO2 elevates the level of intracellular anticancer metabolites in TNBC MDAMB-231 cells, promotes the transformation from glycolysis to aerobic respiration phenotype, and inactivates STAT3 and NF-κB pathyway but activates cAMPpathway TNBC MDA-MB-231 cells, thus affects the malignant biological behaviors of MDA-MB-231 cells.
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@# Objective：To investigate the role and mechanism of chromosomal region maintenance 1 (CRM1) inhibitor sulforaphene (LFS-01) in killing triple negative breast cancer (TNBC) cells by inhibiting signal transducer and activator of transcription 3 (STAT3) signaling pathways. Methods: Whether LFS-01 could combine with the NES pocket of CRM1 was verified by molecular dynamics simulation techniques. The killing activity of LFS-01 on four different breast cancer cell lines was detected by CCK-8 method. TNBC MDA-MB-468 and MDA-MB-231 cells were treated with different concentrations of LFS-01, and the intracellular localization of CRM1 cargo protein STAT3 and protein with NES sequence was detected by immunofluorescence; WB was used to detect the effect of LFS-01 on the expression of STAT-3 signaling pathway and its downstream proteins; WB, cellular immunofluorescence and transmission electron microscopy were adopted to detect the occurrence of autophagy; the effect of LFS-01 on cell cycle and apoptosis was detected by flow cytometry. Results: Molecular dynamics simulations showed that LFS-01 can bind to the NES pocket of CRM1, indicating that it may structurally affect the latter's protein transport function. LFS-01 could specifically kill TNBC MDA-MB-468 and MDA-MB-231 cells. STAT3 and NES-tagged proteins were mainly blocked in the nucleus of TNBC cells after the treatment with 10 μmol/L LFS-01, while they were evenly distributed in the cytoplasm in the control group. The expressions of phosphorylated STAT3 protein, Bcl-xL and Cylin D1 were decreased in MDA-MB-468 and MDA-MB-231 cells with the increase of LFS-01 dose and the prolongation of treatment time; the expression of autophagy marker protein LC3B increased, and highdensity, multi-layered autophagosomes appeared at the same time; cell cycle arrest was observed in S phase and apoptosis rate was significantly increased (P<0.05 or P<0.01). Conclusion: LFS-01 blocks the export of CRM1 carrier protein, thereby inhibiting the activation of STAT3 signaling pathway and promoting autophagy, cell cycle arrest and apoptosis in TNBC MDA-MB-468 and MDAMB-231 cells.
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@#Objective: To investigate the role of exosome (EXO) transporting Let-7a to regulate MYC gene in the malignant biological behaviors of triple negative breast cancer (TNBC) cell, and to explore the underlying mechanism. Methods: After the completion of cell culture, the gene and protein expressions of MYC and Let-7a in TNBC MDA-MB-231cells were detected by qPCR and WB, respectively. Recombinant lenti-virus vector carrying Let-7a and Crisper/Cas-9 system with MYC knockdown were transfected into MDA-MB-231 cells; MTT assay, Transwell assay and Scratch healing assay were performed to examine the proliferation, invasion and migration of MDA-MB-231 cells. Luciferase activity assay was performed to validate the binding between MYC and Let-7a. EXO was isolated and identified by transmission electron microscopy and WB assay in wild-type and Let-7a over-expressed MDA-MB-231 cells, respectively. After co-incubation of two types of EXO and MDA-MB-231 cells, the effects of Let-7a on biological behaviors of MDAMB-231 cells via EXO were detected by qPCR, WB, MTT and Transwell etc. Results: Let-7a was negatively correlated with MYC in breast cancer tissues and cell lines (all P<0.05); MYC promoted while Let-7a inhibited the proliferation, migration and invasion of breast cancer cells (all P<0.01); Let-7a silenced MYC by acting on 3'UTR of MYC gene, thereby reducing the expression of MYC protein (P<0.05); Let-7a was enveloped by EXO and transported to cancer cells, there by inhibiting the proliferation, migration and invasion of MDA-MB-231 cells. Conclusion: EXO some mediated Let-7a silences MYC gene by acting on its 3'UTR region, thus inhibiting the proliferation, migration and invasion of MDA-MB-231 cells.
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@#Objective:To investigate the role of cyclo-oxygenase-2 (COX-2) in breast cancer metastasis and its possible mechanism. Methods: A total of 45 cases of primary breast cancer tissues and brain metastatic breast cancer tissues were collected from patients, who underwent mastectomy in Yunnan Cancer Hospital from October 2015 to April 2018, including 30 cases of primary lesions and 15 cases of brain metastasis. qPCR was used to detect the expression of COX-2 in breast cancer tissues and brain metastatic breast cancer tissues. Recombinant viruses with COX-2 over-expression (LV6-COX2) or COX-2 knockdown (LV3-COX2 shRNA1, LV3-COX2 shRNA2) were transfected into human breast cancer MDA-MB-231 cells; After obtaining the stable expression cell lines, the effect of COX-2 expression on the proliferation of MDA-MB-231 cells was detected by CCK-8, and the effects of COX-2 expression on the migration and invasion of MDA-MB-231 cells were detected by scratch test and Transwell assay, respectively. The mRNAand protein expressions of COX-2 in each group were examined by qPCR and WB, respectively. The effect of COX-2 expression on the expression of EMT-related genes in MDA-MB-231 cells was analyzed by qPCR. Results: The expression of COX-2 in tissues of patients with brain metastases was significantly higher than that in patients with primary breast cancer tissues (P<0.01), and it was correlated with tumor TMN stage in breast cancer patients. MDA-MB-231 cell lines with stable COX-2 over-expression/knockout were successfully constructed. Over-expression of COX-2 promoted the migration and invasion of MDA-MB-231 cells (all P<0.01), and significantly increased the expressions of MMP2, MMP1, N-cadherin and vimentin (all P<0.01), but exerted insignificant effect on cell proliferation. The effect of COX-2 silence exerted the opposite effect and promoted cell proliferation (P<0.05). Conclusion: COX-2 is highly expressed in brain metastatic breast cancer tissues, which may promote the migration and invasion of breast cancer MDA-MB-231 cells by regulating EMT processes.
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@# Objective: To investigate the effect of high expression of TET1 catalytic domain (TET1-CD) gene on the proliferation and migration of breast cancer MDA-MB-231 cells and its underlying mechanism. Methods: MDA-MB-231 cell line with high TET1-CD expression was established by lentiviral transfection. Real-time quantitative PCR was used to detect the mRNA expression of TET1-CD. Transwell assay and cell scratch assay were used to detect cell migration ability, MTT assay and colony formation assay were used to detect cell proliferation capacity. And WB was adopted to detect the expressions of EMT-related proteins (E-cadherin, Vimentin, MMP2) and Wnt, Hedgehog pathway-related proteins in MDA-MB-231 cells. Results: The MDA-MB-231 cell line with high TET1-CD expression was successfully constructed (all P<0.01). TET1-CD over-expression significantly inhibited the proliferation and migration of breast cancer MDA-MB-231 cells (P<0.01); in addition, TET1-CD over-expression increased the expression of E-cadherin, but down-regulated the expressions of Vimentin, MMP2, β-catenin, Gli1, C-myc and CyclinD1 (all P<0.05). Conclusion: TET1-CD may inhibit the proliferation and migration of breast cancer MDA-MB-231 cells by inhibiting the EMT through Wnt and HH signaling pathway.
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Objective:Comprehensive assessment of the chemical composition of sage leaves (Salvia officinalisL.)in order to highlight constituents relevant to the antioxidant, antimicrobial and anticancer potentials, in addition to determining safe dose to facilitate its application in functional foodsand dairy products. Methods:High-Performance Liquid Chromatography (HPLC) was employed to determine constituents such as amino acids, fatty acids and phenolic compounds content. Antioxidant activity was characterized using, ?-diphenyl-?-picrylhydrazyl (DPPH) and reducing power methods. The antimicrobial potentials were examinedagainst nine pathogenic strains. MDA-MB-231 cell line was used to assess anticancer activity. Results:Sage was found to be a good source of calcium, iron andzinc (894.3, 84and 5.5 mg/100g respectively) and vitamins B6 and B12(1.5 and 0.3 mg/100g respectively). PerformedHPLC analysis indicated the rich content of essential amino acids, lysine, phenylalanine andleucine (10.4, 0.7 and 0.45 g/100g),unsaturated fatty acids, Omega 3, 6 and 9(6.46, 4.40 and 3.13g/100g) and phenolic compounds, quercetin and cinnamic (604.8 and 390.4 ?g/mL),which interpreted its high antioxidant powers. Sage revealed antioxidantpotentials with IC50and EC50 reached(27.5 and 239.5 mg/mL respectively), and antimicrobial effect against the examined pathogenic strains with MICs reached 6.25 mm against Staph.aureus, E. coliand Candida albicans, not to mention its anticancer effect as an extra pharmacological feature, when sage performed an anti-proliferative activity with IC50of 300 ?g/mL, against MDA-MB-231 cell line.Conclusion:Obtained results emphasis the sage leaves content of variable nutrients and active compounds that reflected on its vast nutritional and pharmacological potentials such as; antioxidant, antimicrobial and cytotoxic effect against breast MDA-MB-231 cell line, that could nominate it as applicable food bio-preservativein functional foods and dairy products
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OBJECTIVE: To investigate the potential anticancer effect of combretastatin A4 liposomes (SSL-CA4) in destruction of vascular mimicry (VM) in MDA-MB-231 breast cancer cells in vitro. METHODS: The in vitro inhibitory effect and blocking woundhealing effect of SSL-CA4 were investigated. The VM destruction of SSL-CA4 was evaluated in three dimensional matrigel culture model in MDA-MB-231 cells. The in vitro inhibitory test showed that the maxium inhibition ratio of SSL-CA4 on MDA-MB-231 cells was close to 50%. SSL-CA4 blocked the wound-healing of MDA-MB-231 cells, which was similar to free CA4. RESULTS: SSL-CA4 could inhibit the formation of VM in vitro via inhibition on the VM channel indicators including hypoxia-inducible factor (HIF-α), vascular endothelial- cadherin (VE-Cad), and matrix metallopeptidases (MMP-2). The inhibition on indicators of SSL-CA4 was significantly higher than CA4 treatment groups (P <0.05). CONCLUSION: SSL-CA4 has significant anticancer activity via inhibition of VM in MDA-MB-231 cell line.
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Objective@#To investigate the properties of radiation-induced changes of cell cycle and apoptosis in breast cancer cell lines MDA-MB-231 and MCF-7, and to further explore its relationship with radiosensitivity.@*Methods@#The two cell lines MCF-7 and MDA-MB-231 were irradiated with 6 MV X-rays. The cell survival curves were fitted by clonogenic formation assay, then according to the radiosensitivity parameters average lethal dose (D0), quasi-threshold dose (Dq) and survival fraction after 2 Gy irradiation (SF2), the radiosensitivity of the two cell lines was compared. The apoptosis rate and cell cycle distribution of the two cell lines were detected by Hoechst 33342 and PI staining.@*Results@#The survival curve of MDA-MB-231 cell line shifted to the right compared with that of MCF-7 cell line. The values of D0, Dq and SF2 of MDA-MB-231 cell line were higher than those of MCF-7 cell line (1.603±0.023 vs. 1.233±0.027, 1.76±0.04 vs. 1.03±0.10, 0.639 3±0.008 2 vs. 0.398 1±0.018 5, t values were -17.981, -11.745, -20.596, P values were 0.000, 0.003, 0.000). The apoptosis rate of MCF-7 cell line was significantly higher than that of MDA-MB-231 cell line at the doses of 2, 4, 8 Gy irradiated 24 h and at the 12, 48 h after 6 Gy X-irradiation (t values were 4.441, 7.299, 10.499, 6.375, 7.743, P values were 0.011, 0.002, 0.000, 0.003, 0.001). Compared with the non-irradiated group, G2 phase arrest appeared in both cell lines after irradiation. The percentage of the G2/M phase in MDA-MB-231 cell line increased as the time or dosage accumulated. Furthmore, the percentage didn't go down even after 48 hours later. However, the blockage began to gradually release in MCF-7 cell line at the dose of 8 Gy irradiated 24 h and the 48 h after 6 Gy X-irradiation. Followed with that, it turned out the percentage of the G0/G1 phase increased [(65.80±0.56)%, (62.53±0.67)%].@*Conclusions@#6 MV X-irradiation with the doses of 2-8 Gy can induce the cell cycle arrest at G1 and G2 phase in MCF-7 cell line, G2 phase in MDA-MB-231 cell line. Thus more apoptosis appears in MCF-7 cell line, which may cause the difference in radiosensitivity between the two cell lines.
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@#[Abstract] Objective: To establish paclitaxel(PTX)-resistant human triple negative breast cancer cell line and to examine the expression profile of FA-related genes and FANCF, the correlation between the expression of FA-related genes, FANCF and PTX-resistance in breast cancer were further analyzed. Methods: PTX-resistant MDA-MB-231 cell line was established by means of long-term PTX-exposed culture. The sensitivity of the cells to paclitaxel was determined by the CCK8 assay. The cell cycle distribution was examined by flow cytometry after exposure to the paclitaxel. The expression of FA-related gene mRNA and FANCF protein were examined by using real time quantitative PCR and Western blotting. The expression of FANCF in the cells was reduced by RNAi interference technology and the effect of the RNAi was verified. Results: MDA-MB-231/PTX cell showed a 9.9-fold resistance to paclitaxel, indicating that the cell had acquired resistance to PTX. PTX treatment significantly induced G0/G1 arrest and the number of cells in phase S markedly decreased after exposure to PTX. The mRNA and protein expression of FANCF was significantly higher in PTX-resistant cell than that in PTX-sensitve parental cell，Knockdown of FANCF induced apoptosis in MDA-MB-231/PTX cell as well as in parental cell. FANCF knockdown increased the sensitivity of paclitaxel to both MDA-MB-231 and MDA-MB-231/PTX cells (P<0.05 or P<0.01). Conclusion: FANCF played an important role in PTX resistance of the breast cancer cells and FANCF might be a target for therapy aimed at reversing chemoresistance.
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OBJECTIVE To evaluate the effect of Guizhi Fuling Capsule active pharmaceutical ingredient (API) and its fractions on human breast cancer cells proliferation by high-throughput screening assay. METHODS The crude fractions were obtained from the extraction and elution of the API of Guizhi Fuling Capsule, and 929 standard fractions were obtained by the optimal separation conditions. Sulforhodamine B (SRB) method was used to evaluate the effects of the Guizhi Fuling capsule API and 929 kinds of fractions on the proliferation of human breast cancer cells MCF- 7 and MDA- MB- 231. RESULTS The Guizhi Fuling capsule API had a strong ability to inhibit the proliferation of MCF-7 cells at high concentration and the ability to inhibit MDA-MB-231 cells' proliferate at low concentration follow?ing 72 h treatment;some samples of 929 fractions (5 μg·mL-1) was found to have a breast cancer cell growth inhibition rate above 50%, without toxicity on HUVECs proliferation. CONCLUSION The API of Guizhi Fuling capsule had significant cytotoxicity effects on these two human breast cancer cells, with significant concentration- and time-dependent manner.
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Cu(II) and Ni(II) complexes of 2-butyl-4-chloro-5-formylimidazole thiosemicarbazone (L) are synthesized and characterized by using spectroscopic techniques like elemental analysis, FT-IR, mass spectrometry, electronic and EPR spectra. The complexes are found to have characteristic electronic spectra and the geometry of the complexes are identified as octahedron. Both the complexes are found to exhibit similar anti-microbial activity against the gram –ve and gram +ve bacteria. Anti-cancer activity against the cancer cell lines (MDA-MB 231 cell lines) among the compounds studied for % of viability, the inhibition concentration 50 values were shown by Cu(II)-L complex at 80 mg/ml and by Ni(II)-L complex at 100 mg/ml.
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Objective To observe the migration and inhibition mechanism of MicroRNA218-Robo1 pathway for breast cancer.Methods A total of 40 BALB/c-nu/nu female mice were randomly divided into four groups.Each group was transfected over-expression MicroRNA218 MDA-MB-231 breast cancer cells, co-over-expression MicroRNA218 and Robo1 MDA-MB-231 breast cancer cells, knock-down Robo1 MDA-MB-231 breast cancer cells and the control MDA-MB-231 breast cancer cells.The tumor volume was examined every two weeks.Results Tumor volume of MicroRNA218 group was obviously less than control group, tumor volume of Robo1 knock out group was obviously less than common MicroRNA218 high expression and Robo1 group, the difference was statistically significant;MicroRNA218 and Robol knockout group than the control group, the increase in breast cancer cells apoptosis, cell proliferation and angiogenesis is restrained.Conclusions MicroRNA218 inhibited the migration of breast cancer by down-regulating the expression of Robo1.
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Background and purpose:Chemotherapy is the important way of breast cancer treatment, but the drug-resistance has attracted special attention. The emergence of drug resistance is closely related to the abnormal enhancement of DNA-damage repair. Both Kif4A and PARP-1 are important molecules of DNA repair. The research investigated the function of Kif4A in epirubicin up-regulating the activity of PARP-1 in breast cancer cells and possible significance. Methods:Western blot was used to detect the expression of Kif4A and PARP-1 after treatment with epirubicin in MDA-MB-231 and MCF-7 cells; the expression of PARP-1 and its activity were detected after high expression of Kif4A and treatment with epirubicin;FCM was used to detect cell apoptosis after treatment with epirubicin combined with PARP-1 inhibitor 3-ABA. Results:Epirubicin up-regulated PARP-1 activity and induced low expression of Kif4A in breast cancer cells, both of them showed dose-dependent and time-dependent. After high expression of Kif4A, the activity of PARP-1 was inhibited and the apoptosis of cells increased, epirubicin partially reversed the activity of PARP-1 inhibited by high Kif4A expression. Both of epirubicin and 3-ABA induced cell apoptosis, combination of them further increased cell apoptosis compared with alone used (P<0.05). The results also showed the apoptosis rate of MDA-MB-231 cells induced by epirubicin, PARP-1 inhibitor 3-ABA and high expression Kif4A was higher than that of MCF-7 cells (P<0.05). Conclusion:Epirubicin increases the activity of PARP-1 dependent on the low expression of Kif4A in breast cancer cells. Kif4A might become a novel target for overcoming resistance of epirubicin.
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Dietary inorganic sulfur is the minor component in our diet, but some studies suggested that inorganic sulfur is maybe effective to treat cancer related illness. Therefore, this study aims to examine the effects of inorganic sulfur on cell proliferation and gene expression in MDA-MB-231 human breast cancer cells. MDA-MB-231 cells were cultured the absence or presence of various concentrations (12.5, 25, or 50 micromol/L) of inorganic sulfur. Inorganic sulfur significantly decreased proliferation after 72 h of incubation (P < 0.05). The protein expression of ErbB2 and its active form, pErbB2, were significantly reduced at inorganic sulfur concentrations of 50 micromol/L and greater than 25 micromol/L, respectively (P < 0.05). The mRNA expression of ErbB2 was significantly reduced at an inorganic sulfur concentration of 50 micromol/L (P < 0.05). The protein expression of ErbB3 and its active form, pErbB3, and the mRNA expression of ErbB3 were significantly reduced at inorganic sulfur concentrations greater than 25 micromol/L (P < 0.05). The protein and mRNA expression of Akt were significantly reduced at an inorganic sulfur concentration of 50 micromol/L (P < 0.05), but pAkt was not affected by inorganic sulfur treatment. The protein and mRNA expression of Bax were significantly increased with the addition of inorganic sulfur concentration of 50 micromol/L (P < 0.05). In conclusion, cell proliferation was suppressed by inorganic sulfur treatment through the ErbB-Akt pathway in MDA-MB-231 cells.
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Humans , Apoptosis , Breast , Breast Neoplasms , Cell Proliferation , Diet , Gene Expression , ErbB Receptors , RNA, Messenger , SulfurABSTRACT
Objective To study the effects of artesunate(ART)on estrogen receptor negative breast cancer cell line MDA-MB-231 and its mechanisms.Methods Treated with ART for 3 days,MDA-MB-231 cells proliferation was examined by MTT assay.The morphological and uhrastructural changes of MDA-MB-231 cells were observed under microscope and electronic microscope.Immunocytochemistry was used to detect the expression of Bax,mn23,Bcl-2,and P21 WAFl/CIPl.Results ART treatment led to a dose dependent inhibition of MDA-MB-231 cells.ART could change the morphology and ultrastructure of MDA-MB-231 cens.After treatment with ART(2μmol/L)for 72 hours,immunocytochemical staining showed that the expression of Bax,nm23,and P21WAF1/CIP1 was upregulated in comparison to the control group(P0.05).Conclusions ART shows an anti-proliferative effect on MDA-MB-231 cells.The mechanisms may be related to upregnlation of Bax,nm23 and P21WAF1/CIP1 expression.
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In this study, we investigated the effects of the ethanol extract of aerial parts of Raphanus sativus L. (ERL) on breast cancer cell proliferation and gene expression associated with cell proliferation and apoptosis in MDA-MB-231 human breast cancer cells. The MDA-MB-231 cells were cultured in the presence or absence of various concentrations (100, 200, or 300 microg/mL) of ERL. ERL significantly decreased cell proliferation after 48 h of incubation (P < 0.05). The protein and mRNA expression of ErbB2 were decreased significantly in a dose-dependent manner (P < 0.05). The protein expression of ErbB3 was decreased significantly at an ERL concentration of 300 microg/mL (P < 0.05), and mRNA expression of ErbB3 was decreased significantly in a dose-dependent manner (P < 0.05). The protein expression of Akt was decreased significantly at the ERL concentration of 200 microg/mL (P < 0.05), and the protein expression of pAkt was decreased significantly in a dose-dependent manner (P < 0.05). The mRNA expression of Akt was decreased significantly at the ERL concentration of 200 microg/mL ERL (P < 0.05). The protein and mRNA expression of Bax were increased significantly at ERL concentrations of 200 microg/mL or higher (P < 0.05). The protein expression of Bcl2 was increased significantly at ERL concentrations of 100 microg/mL or higher (P < 0.05), and mRNA expression of Bcl2 was increased significantly at an ERL concentration of 300 microg/mL (P < 0.05). In conclusion, we suggest that Raphanus sativus, L. inhibits cell proliferation via the ErbB-Akt pathway in MDA-MB-231 cells.
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Humans , Apoptosis , Breast , Breast Neoplasms , Cell Proliferation , Ethanol , Gene Expression , Raphanus , ErbB Receptors , RNA, MessengerABSTRACT
Objective To investigate four methods for establishing animal models of human breast cancer bone metastasis. Methods Thirty-two female nude mice aged 4-6 weeks were divided randomly into four groups (n=8 in each group). 5×105 MDA-MB-231 cells were injected into the body via the left second mammary fat pads (group A), the tail veins (group B), the left heart ventricles (group C) and the left tibia marrow cavities (group D), respectively. Tumor formations in situ were recorded in group A. Deaths after the injection were recorded. The surviving nude mice 49 days after the injection were subjected to pathological examination to determine bone metastasis. Results The rate of tumor formation in situ of group A was87.5 %(7/8). One mouse in group C died after the injection of MDA-MB-231 cells. The bone metastasis rate in groups A, B, C and D was zero (0/8), 12.5 % (1/8), 71.4 % (5/7) and 100 % (8/8), respectively. There was statistically significant difference in the bone metastasis rate between group A and group C, group A and group D, group B and group C; and group B and group D. Conclusion Injections of tumor cells via the breast fat pads and tail veins were not suitablemethods to establish animal models of human breast cancer bone metastasis. The bone metastasis model could be established efficiently by injecting tumor cells into the left heart ventricles or the bone marrow cavity of nude mice.