Faraday cage-type ECL biosensor for the detection of circulating tumor cell MCF-7.

Herein, a Faraday cage-type electrochemiluminescence biosensor was designed for the detection of human breast cancer cell MCF-7. Two kinds of nanomaterials, Fe3O4-APTs and GO@PTCA-APTs, were synthesized as capture unit and signal unit, respectively. In presence of the target MCF-7, the Faraday cage-type electrochemiluminescence biosensor was constructed by forming a complex "capture unit-MCF-7-signal unit". In this case, lots of electrochemiluminescence signal probes were assembled and could participate in the electrode reaction, achieving a significant increase in sensitivity. In addition, the double aptamer recognition strategy was adopted to improve the capture, enrichment efficiency and detection reliability. Under optimal experimental conditions, the limit of detection was 3 cells/mL. And, the sensor could afford the detection of actual human blood samples, which is the first report on the detection of intact circulating tumor cells by the Faraday cage-type electrochemiluminescence biosensor.

Total Saponins from Paris forrestii Reverse Multidrug Resistance of MCF-7/ADM Cells by Suppression of P-gp via ERK Signaling Pathway.

Our previous study demonstrated that the total saponins from Paris forristii (PCT3) had obvious inhibitory effect on the proliferation of adriamycin-resistant human breast adenocarcinoma cells (MCF-7/ADM), and this effect was significantly stronger than that in parental cells (MCF-7). This study was designed to test the reversal effect of PCT3 on MCF-7/ADM cells and to understand its mechanism of action. Results demonstrated that low cytotoxic concentrations of PCT3 (0.3, 1 and 3 µg/mL) reversed resistance of MCF-7/ADM cells to ADM, cisplatin (DDP) and 5-fluorouracil (5-FU), with reversal fold of 16.4, 19.5 and 31.7 for ADM, 1.6, 1.4 and 1.4 for DDP, 1.7, 1.8 and 5.6 for 5-FU, respectively. Moreover, PCT3 significantly increased the accumulation of ADM and Rhodamine 123 (Rh123) in MCF-7/ADM cells, suggesting that PCT3 may act by affecting the function of drug efflux pump P-glycoprotein (P-gp), which is encoded by MDR1 gene. Both MDR1 gene and P-gp protein expression was downregulated by PCT3 treatment. Further results demonstrated that p38 mitogen-activated protein kinase (MAPK) and extracellular signal-regulated kinase (ERK) pathway was remarkably activated in MCF-7/ADM cells, inhibition of p38 or ERK attenuated P-gp expression. While, only the phosphorylation level of ERK was downregulated by PCT3, indicating that PCT3 sensitized P-gp overexpressed MCF-7/ADM cells to ADM via inhibition of ERK signaling pathway.

Griffipavixanthone induces apoptosis of human breast cancer MCF-7 cells in vitro.

BACKGROUND: Griffipavixanthone (GPX) is a compound extracted from Garcinia oblongifolia Champ. But, no research has yet been done about the effect of GPX on breast cancer. METHODS: We evaluated the proliferation of human breast cancer cells by CCK-8 assay and apoptosis by Annexin V (AV)-FITC and PI double staining. We used transwell assay to indicate the invasion and migration of MCF-7. To explore the molecular mechanism of GPX, we detected the mRNA and protein expression using qRT-PCR and Western blot. RESULTS: In this study, we evaluated if GPX could inhibit the proliferation of human breast cancer cell MCF-7 and T-47D with IC50 value of 9.64 ± 0.12 µM and 10.2 1 ± 0.38 µM at 48 h. And the IC50 value of MCF-10A is 32.11 ± 0.21 µM, which showed GPX had a tiny side effect for normal breast cells. Annexin V (AV)-FITC and PI double staining demonstrated firmly the apoptosis of MCF-7 resulting from GPX. Transwell assay indicated that GPX inhibited the invasion and migration of MCF-7. In addition, we found GPX cleaved caspase-8/9 and PARP, which play important roles in apoptotic pathway. Furthermore, through the Western blot assay, GPX increased the level of pro-apoptosis protein Bax and cytochrome C. On the contrary, GPX decreased the level of anti-apoptosis protein Bcl-2. Moreover, GPX increased the mRNA and protein expression level of p53 and its target genes, which indicated that GPX induced MCF-7 cell apoptosis by up-regulating p53 and Bax expression while suppressing Bcl-2 expression. CONCLUSION: All the results showed that GPX induces MCF-7 cell apoptosis and could be considered as a potential drug for breast cancer.

Effect of dihydromyricetin on proliferation and apoptosis of breast cancer MCF-7 cells / 国际检验医学杂志

Objective To investigate the effect of dihydromyricetin on proliferation and apoptosis of breast cancer MCF-7 cells.Methods From March 2014 to February 2015,breast cancer MCF-7 cells were treated with 99% pure DMY as an inhibitor.MTT assay,flow cytometry and immunocytochemistry were used to analyze the proliferation,apoptosis and protein expression of breast cancer cell MCF-7.Results When the DMY concentration was higher than 20 μg/mL,the inhibitory effect appeared,but not good.When 40 and 80 μg/mL DMY were used,the proliferation of MCF-6 cells were significantly inhibited,and have different degrees of sensitivity to it.When DMY was 80 μg/mL,the IC50 was 226.9 μg/mL.The inhibition rate and IC50 were compared with 0 μg/mL DMY,there was significant difference(P 50%,especially in DMY with 80 μg/mL,the positive rate was 10.00%.Compared with 0 μg/mL DMY,the difference was significant(P<0.05).Conclusion The use of dihydromyricetin in breast cancer patients can effectively inhibit the rapid increase of cancer cells,accelerate apoptosis,slow down the patient′s condition,the effect is outstanding.

Effect of D-α-tocopherol polyethylene glycol 1000 succinate on inhibition of MCF-7 cell proliferation by baohuoside I / 中草药

Objective: To investigate the effect of D-α-tocopherol polyethylene glycol 1000 succinate (TPGS) on the inhibition of proliferation of breast cancer cells MCF-7 by baohuoside I. Methods: The cytotoxicity of baohuoside I to MCF-7 cells was determined by MTT assay, the cellular uptake of baohuoside I was detected by fluorescence microscopy, and the intracellular baohuoside I was determined by HPLC. Results: The effect of baohuoside I on the inhibition of MCF-7 cell proliferation was enhanced in the presence of TPGS, especially on lower concentration. The uptake rates of MCF-7 within 2 h were 29.51%, 38.12%, and 40.37%, when the proportions of baohuosaide I and TPGS were 1:1, 1:2, and 1:4, respectively. The ratios were increased by 27.92%, 65.24%, and 74.99% compared with those using baohuoside I only. Conclusion: TPGS can increase the uptake rate of baohuoside I in MCF-7 cells and enhance the inhibition of MCF-7 cell proliferation.

DLJ14, a novel chemo-sensitization agent, enhances therapeutic effects of adriamycin against MCF-7/A cells both in vitro and in vivo.

OBJECTIVES: We investigated the chemo-sensitization of a ligustrazine derivate, (E)-2-(2, 4-dimethoxystyryl)-3, 5, 6-trimethylpyrazine (DLJ14) on Adriamycin (Adr, Wanle, Shenzhen, China)-resistant human breast cancer (MCF-7/A) cells both in vivo and in vitro. METHODS: The antitumour effects of DLJ14 and Adr was observed in MCF-7/A cells by 3-(4,5-dimethyl-thiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay in vitro and was evaluated by MCF-7/A xenografts in nude mice. The intracellular Adr accumulation was assessed by mean fluorescence intensity of Adr. The messenger RNA level of glutathione (GSH) S-transferase (GST)π in MCF-7/A cells was determined by real-time reverse transcription PCR assay. The expression of GSTπ, c-jun NH2 -terminal kinase (JNK) and phosphor-JNK (p-JNK) was detected by Western blotting method. KEY FINDINGS: The MTT results showed that DLJ14 exhibited a weak inhibition on proliferation of both MCF-7 and MCF-7/A cells, in contrast with the strong inhibition of verapamil. When DLJ14 is combined with Adr, the inhibitory effect on MCF-7/A cells and MCF-7/A xenografts was enhanced significantly through increasing intracellular accumulation of Adr by inhibition of GSH level and the activity of GSH peroxidase and GST. Moreover, DLJ14 could downregulate the expression of GSTπ and increase the expression of JNK and p-JNK in MCF-7/A cells or in xenografts. CONCLUSION: DLJ14 is a promising chemo-sensitization candidate for the reversal of multidrug resistance in cancers.

Effects of estradiol plus testosterone on the proliferation and apoptosis of breast cancer cell line MCF-7 / 肿瘤

Objective: To study the effects of estradiol plus testosterone on the proliferation and apoptosis of breast cancer cell line MCF-7, and to elucidate the possible mechanism. Methods: The growth of MCF-7 cells treated with estradiol (10-10 mol/L) or testosterone (10-5, 10 -7, 10-9 and 10-11 mol/L) alone or in combination for 24, 48 and 72 h was detected by MTT assay. The cell cycle distribution and the apoptosis rate of MCF-7 cells were determined by flow cytometry (FCM). The expression levels of cyclinD1 and androgen receptor (AR) proteins were examined by FCM. Results: The proliferation ability of MCF-7 cells was elevated after treatment with estradiol, and this effect could be inhibited by a higher concentration of testosterone (10-5 mol/L) while improved by a lower concentration of testosterone (10-9 mol/L). After treatment with estradiol plus testosterone (10-5 mol/L) for 48 h, the G1/S phase transition of MCF-7 cells was accelerated, and the apoptosis rate was increased; the expression level of cyclinD1 protein was increased while no change of AR protein expression was observed. Conclusion: Estradiol combined with testosterone of high concentration can inhibit the proliferation of MCF-7 cells and improve the apoptosis. This effect may be associated with the up-regulation of cyclinD1 expression. Copyright© 2011 by the Editorial Board of Tumor.

Effect of fucoidan on proliferation and apoptosis of human breast carcinoma cell line MCF-7 / 肿瘤

Objective: To investigate the effects of fucoidan on proliferation and apoptosis of breast carcinoma cell line MCF-7 and the underlying mechanism. Methods: Cell viability was measured by MTF assay. Apoptosis was detected by Hoechst 33258 staining and agarose gel electrophoresis. The mRNA and protein expression of bcl-2 and bax were examined by RT-PCR and Western blotting, respectively. Results: Fucoidan effectively inhibited the proliferation of MCF-7 cells in a concentration-dependent manner (P <0.01). Fucoidan induced significant apoptosis of MCF-7 cells characterized by fragmented DNA ladder. Fucoidan down-regulated the expression of bcl-2 and up-regulated bax at mRNA and protein levels. The ratio of bcl-2 to bax decreased with the increase in the fucoidan concentrations (P < 0.05). Conclusion: Fucoidan inhibites the proliferation and induced the apoptosis of MCF-7 cells. The underlying mechanism may be related with down-regulation of anti-apoptotic protein bcl-2 and up-regulation of apoptotic protein bax.

Effects of MST1 on cell proliferation and apoptosis of human breast carcinoma cell line MCF-7 / 肿瘤

Objective: To elucidate effects of mammalian sterile 20-like kinase 1 (MST1) gene on cell proliferation and apoptosis of human breast carcinoma cell line MCF-7. Methods: This study constructed plasmid pCMV-FLAG-MST1 and transfected it into human breast carcinoma cell line MCF-7 via mediation by Lipofectamine 2000. We detected the efficiency of transfection by Western blotting. The effect of MST1 on cell proliferation was measured by MTT assay at 12, 24, 36, and 48 h after transfection. The effect of MST1 on cell proliferation was also determined by BrdU incorporation assay at 36 h after transfection. Cisplatin was added into cultured cells at 36 h to induce apoptosis. Forteen hours later cell apoptosis was detected by Annexin V staining. Results: This study successfully constructed plasmid pCMV-FLAG-MST1. The MTT and BrdU incorporation assay showed that the cell proliferation was significantly inhibited after transfection of pCMV-FLAG-MST1. Annexin V staining demonstrated that cell apoptotic rate was relatively higher after overexpression of MST1. Conclusion: In summary, overexpression of MST1 decreases cell proliferation and induced apoptosis of human breast carcinoma cell line MCF-7.

Effects of aspirin and celecoxib on expressions of COX-2 and VEGF in breast cancer cells MCF-7 / 吉林大学学报(医学版)

0.05),the inhibitory rates in the other groups were increased compared with control group(P0.05),the expressions of COX-2 and VEGF in MCF-7 cells in the other groups were lower than that in negative control group(P

Inhibitory Effect of K_6[P_2W_(18)O_(62)]?14H_2O on Human Breast Tumor Cell MCF-7 Growth in Vitro / 中国药房

OBJECTIVE: To study the in vitro anti-tumor activity of tungstophosphate compound(K6[P2W18O62]?14H2O) with Dawson structure.METHODS: MCF-7 cells were treated with different concentration of K6[P2W18O62]?14H2O,then its IC50 was measured by MTT assay;flow cytometry was applied to analyze cell cycle and apoptosis by computing the ratio of MCF-7 cells in different phase and its apoptotic ratio.Cell growth and viability were measured by trypan blue staining and cell counting by computing the ratio of living cells.The synergetic anti-tumor activity of tungstophosphate compound(K6[P2W18O62]?14H2O) and pharmorubicin was studied.RESULTS: K6[P2W18O62]?14H2O inhibited MCF-7 cell proliferation with IC50 of(33.7?3.2) ?mol?L-1,it reduced the proportions of MCF-7 cells at S phase and G1 phase,with the apoptotic rate at 3.7%～29.2% within 48 h.The percentage of living cells was 95.37%～76.78% at 48 h.As compared with pharmorubicin alone,the addition of K6[P2W18O62]?14H2O to pharmorubicin showed a significantly higher inhibitory action on proliferation of MCF-7 cells(P