In vitro evaluation for estrogenic mechanisms of the disinfectant benzalkonium chloride as an emerging contaminant

This study aimed to evaluate in vitro the possible mechanisms underlying the estrogenic potential of benzalkonium chloride (BAC) as a disinfectant emerging contaminant. Effects of BAC at the environmentally-relevant concentrations on estrogen synthesis and estrogen receptor (ER) signaling were assessed using the H295R steroidogenesis assay and the MCF-7 proliferation assay, respectively. Results showed that exposure to BAC at concentrations of 1.0-1.5 mg/L for 48 h significantly increased estradiol production of H295R cells in a concentration-dependent manner. Transcription of steroidogenic genes 3β‐HSD2, 17β‐HSD1, 17β‐HSD4, and CYP19A were significantly enhanced by BAC. In ER-positive MCF-7 cells, exposure to 0.5-1.5 mg/L BAC for 48 h significantly promoted cell proliferation and increased the expressions of ERα and G-protein coupled estrogen receptor 1. Flow cytometry analysis showed that 0.5-1.5 mg/L BAC significantly decreased the percentage of cells in G0/G1 phase, increased the percentage in S phase, and BAC at concentrations of 1.0 and 1.5 mg/L increased the G2/M phase cells. Findings of the study suggested that BAC at environmentally-relevant concentrations might act as a xenoestrogen through its inhibitory effect on steroidogenesis and ER-mediated mechanism.

Expression of Tspan29 in breast cancer tissues and its effect on malignant biological behaviors of MCF-7 and MDA-MB-231 cells / 中国肿瘤生物治疗杂志

@#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. ··

MiR-142-3p enhances chemosensitivity of breast cancer cells and inhibits autophagy by targeting HMGB1

MiR-142-3p has been reported to act as a tumor suppressor in breast cancer. However, the regulatory effect of miR-142-3p on drug resistance of breast cancer cells and its underlying mechanism remain unknown. Here, we found that miR-142-3p was significantly downregulated in the doxorubicin (DOX)-resistant MCF-7 cell line (MCF-7/DOX). MiR-142-3p overexpression increased DOX sensitivity and enhanced DOX-induced apoptosis in breast cancer cells. High-mobility group box 1 (HMGB1) is a direct functional target of miR-142-3p in breast cancer cells and miR-142-3p negatively regulated HMGB1 expression. Moreover, overexpression of HMGB1 dramatically reversed the promotion of apoptosis and inhibition of autophagy mediated by miR-142-3p up-regulation. In conclusion, miR-142-3p overexpression may inhibit autophagy and promote the drug sensitivity of breast cancer cells to DOX by targeting HMGB1. The miR-142-3p/HMGB1 axis might be a novel target to regulate the drug resistance of breast cancer patients.

Mechanism of microRAN-145 on proliferation and apoptosis of breast cancer cell line MCF-7 / 解剖学报

Objective To explore the mechanism of microRNA-145 (miR-145) involved in proliferation and apoptosis of breast cancer MCF-7 cells. Methods The immortalized breast cancer cell line MCF-7 cells were cultured in vitro and transfected with miR-145. Real-time PCR and Western blotting were used to detect mRNA and protein levels, respectively. The proliferation level of each group was detected by cell counting kit-8 (CCK-8) method , and the apoptosis level of MCF-7 cells in different treatment groups was detected by flow cytometer. Results Real-time PCR result showed that miR-145 did not affect Caspase-3, proliferating cell nuclear antigen (PCNA) and B cell lymphoma factor 2 (Bcl-2) mRNA levels in MCF-7 cells. Western blotting analysis showed that compared with the control group, transfection of miR-145 for 96 hours significantly increased the expression of Caspase-3 and inhibited the expression of PCNA and Bcl-2. The result of CCK-8 assay showed that the proliferation rate of MCF-7 cells was decreased after overexpression of miR-145 for 72 hours and 96 hours (P<0. 05). The result of flow cytometer showed that the apoptosis rate of MCF-7 cells in overexpression group was significantly higher than that in the control group (P<0. 05). Conclusion MiR-145 can inhibit the proliferation and promote the apoptosis of MCF-7 cells by down-regulating PCNA and Bcl-2 and up-regulating the expression of Caspase-3, which may be a new target for breast cancer treatment.

lncRNA LINC00969 inhibits breast cancer cell proliferation and migration / 中国肿瘤生物治疗杂志

@#[Abstract] Objective: To investigate the effect of long non-coding LINC00969 on proliferation and invasion of breast cancer cell. Methods: Real-time Quantitative polymerase chain reaction (qPCR) was used to detect differential expression of LINC00969 in five breast cancer cell lines (MCF-7, BT-20, MAD-MB-231, ZR-75-1, and SKBR3) , normal breast cells MCF-10A, and in 42 cases breast cancer tissues and adjacent tissues. TMCF-7 cells were transfected with LINC00969 plasmid and empty vector vector vector. The transfection efficiency was verified by qPCR. CCK-8, plate cloning and edu assay were used to detect cell proliferation. Flow cytometry was used to detect cell cycle. Western blotting was used to detect PCNA, CyclinD1, MMP2 and MMP9. Scratch repair test and Transwell test were used to detect cell migration and invasion. Results：Compared with the adjacent tissues, LINC00969 expression in breast cancer tissues was significantly decreased (P<0.05); compared with breast cancer cells MCF-10A, LINC00969 expression in five breast cancer cells was significantly decreased (P<0.05), and the lowest was in MCF-7 cells; overexpression of LINC00969 significantly inhibited the proliferation, colony formation and DNA synthesis of MCF-7 cells (all P<0.05), making MCF-7 cell cycle clear.The ability of wound healing, migration and invasion of the cells were significantly reduced (P<0.05). Overexpression of LINC00969 significantly inhibited the expression of PCNA, cyclinD1, MMP2 and MMP9 (all P<0.05). Conclusion: LINC00969 is low expressed in breast cancer. Overexpression of LINC00969 can inhibit proliferation and migration of breast cancer cell,the mechanism may be related to the abnormal expression of cell cycle and migration related proteins.

Anti-tumor effect of new LL-37 hybrid antimicrobial peptide on breast cancer MCF-7 cells / 中国肿瘤生物治疗杂志

@#[Abstract] Objective:To investigate the anti-tumor effect and mechanism of new LL-37 hybrid peptide on breast cancer MCF-7 cells. Methods: Human antimicrobial peptide LL-37 and human neutrophil peptide 1(HNP-1) were screened by using of Antimicrobial Peptides Database (http:// aps.unmc.edu/AP/main.php). The new LL-37 hybrid peptide was synthesized by integrating the active fragments, which were selected by bioinformatics analysis. The breast cancer MCF-7 cells and human normal breast MCF10A cells were treated with the new LL-37 hybrid peptides (0~70 μmol/L). Cell viability was monitored by CCK-8 assay and the affinity of the new LL-37 hybrid peptide with MCF-7 cells was observed using confocal laser scanning microscope (CLSM). The effects of LL-37 and caspase inhibitor on apoptosis and cell cycle of MCF-7 cells were measured by FCM (flow cytometry). Results: The new LL-37 hybrid peptide, as an amphiphilic cationic polypeptide, could selectively inhibit the proliferation of breast cancer MCF-7 cells ( P <0.05) with an IC50of 58.34 μmol/L, but exerted no significant effect on normal breast MCF10A cells. LL-37 peptide had high affinity with MCF-7 cells, which could cause S-stage stagnation and significantly increased early apoptosis ( P <0.01); however, the cell cycle block and apoptosis were significantly attenuated after the treatment of caspase inhibitor ( P <0.01). Conclusion: The new LL-37 hybrid peptide has anti-tumor activity on breast cancer MCF-7 cells, and could induce MCF-7 cells apoptosis possibly by arresting cell cycle via the caspase-dependent signaling pathway.

miR-1297 promotes malignant biological behaviors of breast cancer MCF-7 cells by down-regulating TET3 / 中国肿瘤生物治疗杂志

@# Objective: To investigate the regulatory effect of miR-1297 on the malignant biological behaviors of breast cancer cells and its underlying mechanism. Methods: Twenty pairs of breast cancer tissues and para-cancer tissues resected at the Department of Thyroid and Breast Surgery of Leshan People′ s Hospital from May 2016 to May 2018, as well as breast cancer cell lines MCF-7, SW626, HCC1937 and human breast epithelial MCF-10A cells were collected for this study. qPCR was performed to evaluate the expression of miR-1297 in breast cancer tissues and cell lines. The experimental cells were divided into control group, miR-1297 inhibitor group; TET3 over-expression group and simultaneous over-expression of TET3 and miR-1297 group. CCK-8 assay was used to detect the cell proliferation of MCF-7 cells; Transwell assay was carried out to detect the migration and invasion of MCF-7 cells; and WB was used to measure the expressions of TET3 and EMT related proteins (E-cadherin, N-cadherin and vimentin). Dual luciferase reporter gene assay was used to verify the relationship between miR-1297 and TET3. Results: miR-1297 was up-regulated in both breast cancer tissues and cell lines (P<0.01 or P<0.05). Knockdown of miR-1297 dramatically repressed the proliferation, migration, invasion and EMT of MCF-7 cells (P<0.01 or P<0.05). Over-expression of TET3 significantly up-regulated the expression of TET3 in MCF-7 cells (P<0.05). Simultaneous over-expression of TET3 and miR-1297 could reverse the expression level of TET3 in MCF-7 cells and the inhibitory effect of TET3 on the proliferation, migration, invasion and EMT of MCF-7 cells. Dual luciferase reporter gene assay results showed that miR-1297 targetedly bound to the 3' UTR of TET3. Further experiment results demonstrated that miR-1297 targetedly down-regulated TET3 and promoted the malignant biological behaviors of MCF-7 cells. Conclusion: miR-1297 is up-regulated in breast cancer tissues and cells; it promotes the malignant biological behaviors such as proliferation, migration, invasion and EMT through targetedly down-regulating the expression of TET3.

Effect of Tryptanthrin on Proliferation of Human Breast Cancer MCF-7 Cells Via MAPK Signaling Pathway / 中国药学杂志

OBJECTIVE: To investigate the effect of tryptanthrin on human breast cancer MCF-7 proliferation and MAPK signaling pathway. METHODS: The human breast cancer cell MCF-7 was cultured in vitro, different concentrations of 0.1, 0.5, 1, 5, 10, 25, 50, 100 μmol•L-1 of tryptanthrin and ERK, p38MAPK and JNK pathway inhibitors were used to treat MCF-7 cell for 24, 48 or 72 h. MTT assay was used to detect cell proliferation activity. The expression level of ERK, p-ERK, p38, p-p38, JNK and p-JNK were measured by Western blot method. RESULTS: Low, medium and high doses of tryptanthrin significantly inhibited the proliferation of MCF-7 cells. When MCF-7 cells were treated with different concentrations of tryptanthrin for 24, 48, 72 h, the inhibition rate of cell proliferation was correlated with the drug concentration (r=0.904, r=0.793, r=0.770, P<0.05). The 25 μmol•L-1 of tryptanthrin significantly inhibits the proliferation of MCF-7 cells for 24 h treatment(P<0.01). The cell proliferation activity of tryptanthrin combined with ERK inhibitor group, tryptanthrin combined with p38 MAPK inhibitor group, tryptanthrin combined with JNK inhibitor group was significantly different from that of tryptanthrin group(P<0.01 or P<0.001); MCF-7 cells was treated with 6.25, 12.5, 25 μmol•L-1 of tryptanthrin, the expression of p-ERK and p-JNK increased significantly(P<0.05 or P<0.01); MCF-7 cells was treated with 25 μmol•L-1 of tryptanthrin, the expression of p-p38 MAPK increased significantly(P<0.05); the expression of ERK, p38 MAPK and JNK did not change significantly. CONCLUSION: Tryptanthrin can inhibit the proliferative activity of MCF-7 cells, and its mechanism may be closely related to the activation of MAPK signaling pathway.

Effect of Bak gene overexpression on proliferation of breast cancer MCF-7 cells and sensitivity to paclitaxel / 解剖学报

Objective: To investigate the effect of Bak gene overexpression on proliferation of breast cancer MCF-7 cells and sensitivity to paclitaxel. Methods: Western blotting and Real-time PCR were used to detect the expression of Bak protein in breast cancer MCF-7 cells before and after transfection. The effects of Bak gene overexpression and paclitaxel treatment for 24 hours, 48 hours and 72 hours on MCF-7 cell proliferation and cell cycle were detected by cell counting kit-8(CCK-8) and flow cytometry. Results: The results of Western blotting and Real-time PCR showed that the expression of Bak protein increased significantly after transfection. The results of Western blotting and Real-time PCR showed that the expression of Bak protein increased significantly after transfection, and the mRNA expression of MCF-7-Bak group was 2.15±0.07, which was significantly higher than that of MCF-7-NC group, which was 1.03±0.04(r= 13.412, P <0.05). After 48 hours, 72 hours and 96 hours of transfection, the proliferation rates of MCF-7-Bak cells were (0.31 ± 0.03)%, (0.37±0.03)%, (0.47±0.04)%, respectively, which were lower than that of MCF-7 NC group (0.40± 0.03)%, (0.48±0.04)%, (0.61±0.06)%, and the difference was statistically significant (t48 = 2.145, t72 = 3.164, t96 = 5.487, P<0.05). The number of G2 phase cells in MCF-7 Bak group were (26.84±2.69)% significantly higher than that in MCF-7 NC group (16.02±1.61)% ({=12.887, P<0.05). After 24 hours, 48 hours, and 72 hours of paclitaxel treatment, the cell inhibition rate of MCF-7-Bak group was (35.98±4.00)%, (54.66±5.50) %, (80.11 ± 8.00) %, respectively, which were higher than that of MCF-7 NC group (24.12±2.40)%, (40.12±4.00)%, (61.09±6.09)%. And the difference was statistically significant (t24 = 8.456, t48 = 10.547, t72 = 13.442, P<0.05). After 24 hours of paclitaxel treatment, the number of G2 phase cells in MCF-7 Bak group was (73.01 ±7.02)% higher than that in MCF-7 NC group (63.84±6.68) %(t = 9.736, P<0.05). Conclusion: Up-regulate of Bak gene expression can inhibit the proliferation of breast cancer MCF-7 cells, up-regulating the proportion of G0/G1 phase, and enhance the sensitivity of paclitaxel.

Antiproliferative potential of solidagenone isolated of Solidago chilensis

ABSTRACT Plants are considered among the main sources of biologically active chemicals. The species Solidago chilensis Meyen, Asteraceae, is native to the southern parts of South America, where the aerial parts of the plant are commonly used for the treatment of inflammatory conditions. However, the effects of S. chilensis on human cancer cells remain to be elucidated. In this study, we evaluated the antiproliferative effects of the hydroalcoholic and dichloromethane extracts of S. chilensis, as well as their chemical constituents quercitrin and solidagenone against the five human tumor cell lines in vitro. The dichloromethane extract showed a promisor antiproliferative effects in vitro, especially against glioma cell line. Besides, the hydroalcoholic extract and quercitrin were inactive. The diterpene solidagenone showed highly potent antiproliferative effects against breast (MCF-7), kidney (786-0), and prostate cancer (PC-3) cells (total growth inhibition: TGI < 6.25 µg/ml). Solidagenone meets the theoretical physico-chemical criteria for bioavailability of drugs, according to the "Rule of Five" and, by theorical studies, the observed biological effects were probably related to the interaction of the molecule with nuclear receptors and as an enzymatic inhibitor. This study contributes to chemical study and to the identification of antiproliferative molecules in S. chilensis.

Determination of the binding site of hypoxia inducible factor-2a to the MMP-2 promoter / 临床与实验病理学杂志

Purpose To identify the binding of hypoxia in-ducible factor-2a (HIF-2 a) to the hypoxia-response element(HRE) of the matrixmetallo proteinases-2 (MMP-2) gene promoter region and clear the binding site. Methods Electro-phoretic mobility shift assay (EMSA) was used to identify the binding of HIF-2 a to HRE of the MMP-2 gene promoter region in vitro. At the same time, the chromatin immunoprecipitation assay (CHIP) was used to further determine the binding site. Results Successful prediction of two potential HIF-2a binding sites of MMP-2 the promoter region, which were-217～-204 and-1 029 ～-1 007, respectively. Probe test shows that the marked efficiency of sense chain and antisense chain was above 50％, and they could be used for EMSA-electrophoretic mobility shift assay. The results of EMSA showed that there was a binding site of HIF-2 a sense chain and antisense chain moter region int-217～-204. The results of chromatin immuno-precipitation showed that in the experimental group and control group an about 250 bp fragment in MMP-2 promoter containing HRE region was amplified, suggesting that the protein of HIF-2a binded to the HRE in MMP-2 promoter region in vivo. Conclusion HIF-2 a in MMP-2 promoter regionne promoter region in vitro and in vivo.

Effect of radiation-induction on cell cycle and apoptosis in breast cancer cell lines MDA-MB-231 and MCF-7 / 肿瘤研究与临床

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.

Expression of galectin-3 in human breast cancer tissues and its effect on malignant biological behavior of MCF-7 cells / 中国肿瘤生物治疗杂志

@# Objective: To investigate the expression of galectin-3 protein in human breast cancer tissues and the effect of silencing galectin-3 gene on the migration, invasion and apoptosis of human breast cancer MCF-7 cells. Methods: The relative expression of galectin-3 protein in 15 cases of breast cancer tissues and corresponding para-cancerous tissues were detected by Western blotting; The expression of galectin-3 protein in paraffin sections of 100 cases of breast cancer tissues were detected by immunohistochemistry, and the correlation between galectin-3 expression and the clinicopathological characteristics of breast cancer patients was also analyzed. Galectin-3 siRNA were transfected into human breast cancer MCF-7 cells by liposome, then Real-time PCR and Western blotting were used to detect the mRNA and protein expression of galectin-3. The effect of galectin-3 gene silencing on cell migration and invasion ability of MCF-7 cells were detected by Transwell method. The effect of galectin-3 gene silencing on apoptosis of MCF-7 cells were detected by flow cytometry. Results: Western blotting detection showed that the relative expression of galectin-3 protein in breast cancer tissues were significantly higher than that in para-cancerous tissues (P<0.05); Immunohistochemistry detection showed that the positive expression rate of galectin-3 protein in breast cancer tissues was 67.00%, the positive expression rates in the lymph node metastasis, hormone receptor (ER, PR) negative groups were significantly higher (P<0.05), and the positive expression rate of galectin-3 protein were increased with the increase of TNM stage and histological grade (P<0.05); Galectin-3 siRNA transfection could significantly reduce the mRNAand protein expression of galectin-3 in MCF-7 cells (P<0.05), and reduce the invasion and migration ability but significantly improve the rate of apoptosis of MCF-7 cells (P<0.05). Conclusion: Galectin-3 is highly expressed in breast cancer tissues, and its silence can inhibit the invasion and metastasis of MCF-7 cells and induce apoptosis of MCF-7 cells. Galectin-3 can be used as a new target for biological therapy of breast cancer.

Expression of lncRNA RP3-340N1.2 in breast cancer tissues and its effect on proliferation and migration of MCF-7 cells / 中国肿瘤生物治疗杂志

@#Objective: To observe the expression of long-chain non-coding RNA (lncRNA) RP3-340N1.2 in breast cancer tissues and its effect on proliferation and migration of breast cancer MCF-7 cells, and to explore the possible mechanism. Methods: 13 pairs of breast cancer tissues and adjacent tissues from breast cancer patients, who underwent radical surgery at the Cancer Center of theAffiliated People’s Hospital of Hubei University of Medicine from Jan. 2017 to Sep. 2017, were collected for this study. qRT-PCR was used to detect the differential expression of RP3-340N1.2 in collected tissue samples and breast cancer cell lines and normal breast epithelial cell line. RP3-340N1.2 plasmid (experimental group) and the negative control plasmid (control group) were transfected into breast cancer MCF-7 cells using Lipofectamine 3000. Cell counting (CCK-8) and Transwell migration assay were used to examine the effect of RP3-340N1.2 over-expression on proliferation and migration of MCF7 cells, the effect of RP3-340N1.2 over-expression on the mRNA expression of miR-134-5p and OPCML was detected by qRT-PCR, and Western blotting was used to detect the expression of OPCML protein. Results: The expression of RP3-340N1.2 in breast cancer tissues was significantly lower than that in adjacent tissues ( P <0.01), and the expression of RP3-340N1.2 in breast cancer cell lines was significantly lower than that in normal breast epithelial cells ( P < 0.01). Up-regulation of RP3-340N1.2 decreased the proliferation and migration of MCF7 cells (all P <0.05). After over-expression of RP3-340N1.2 in MCF7 cells, the expression of miR-134-5p obviously decreased ( P <0.01); moreover, the mRNA and protein expressions of OPCML significantly increased ( P <0.01) while the expressions of cell cycle regulatory proteins (CDK4, Cyclin D2) and cell migration regulatory proteins (Vimentin and N-cadherin) decreased significantly (all P <0.01). Conclusion: RP3-340N1.2 is low expressed in breast cancer tissues and cell lines. Up-regulation of RP3-340N1.2 expression can lead to decreased expression of miR-1345p and increased expression of OPCML gene, thereby inhibiting the proliferation and migration of breast cancer cells.

Effect of Saposhnikovan Sulphate on the Recruitment and Re-education of Macrophages by MCF-7 Cells In Vitro through CCL3 / 中国药学杂志

OBJECTIVE: To observe the inhibition of saposhnikovan sulphate obtained by ultrasonic extraction(S-USPS) on human leukemia MCF-7 cells in vitro and the effect of S-USPS on the recruitment and re-education of macrophages by MCF-7 cells. METHODS: S-USPS was obtained from saposknikovan(SPS) by the method of chlorosulfonic acid-pyridine complex. The inhibition of S-USPS and USPS on proliferation of MCF-7 cells was measured by MTT. THP-1-derived macrophages were differentiated in vitro. The effect of S-USPS on the recruitment of macrophages by MCF-7 cells was detected by transwell migration assay. The effects of S-USPS on the expression and secretion of chemokine were detected by qRT-PCR and ELISA. The effects of S-USPS and CCL3 on the expression of inflammatory cytokines were detected by qRT-PCR. RESULTS: The inhibitory rate of S-USPS on the proliferation of MCF-7 was dose-dependent and time-dependent and was higher than that of USPS(P<0.05).S-USPS increased the transcription and secretion of CCL3 by MCF-7 cells. There were higher expressions of IL-1β, IL-6, and TNF-α mRNA and lower expressions of TGF-β, IL-10 mRNA of macrophages in the S-USPS group than in the control group and MCF-7 supernatant group(P<0.05). The promotion of S-USPS on the expressions of IL-1β and TNF-α was inhibited by CCL3 neutralizing antibody(P<0.05). CONCLUSION: There are significant evidences that the saposhnikovan sulphate effectively promotes the recruitment of macrophages by MCF-7 cells in vitro, and re-educates their secretion profile toward M1-type.

Effects of cadmium on the expression of estrogen receptor and miRNA in MCF-7 cells / 中国职业医学

OBJECTIVE: To study the effects of cadmium on the expression of estrogen receptor( ER) and miRAN-155,miRAN-200 c in human breast cancer MCF-7 cells. METHODS: MCF-7cells in logarithmic growth phase were randomly divided into fulvestrant( ICI182780,ICI) group and non-ICI group. The non-ICI group was treated with cadmium chloride(Cd Cl2) at the final concentrations of 0. 0,2. 5,5. 0 and 10. 0 μmol/L for 24 hours. The ICI group was pretreated at a concentration of 1. 0 μmol/L for 12 hours,and then treated with Cd Cl2 at the final concentrations 0. 0,2. 5,5. 0 and 10. 0μmol/L for 24 hours. The cell proliferation activity was measured by methyl thiazolyl tetrazolium assay. Flow cytometry was used to measured cell apoptosis. Western blot was applied to measure the relative expression of ERα and ERβ protein,and the relative expression of miRNA-155 and miRNA-200 c were detected by real-time fluorescence quantitative polymerase chain reaction. RESULTS: The proliferation rates of MCF-7 cells in 2. 5,5. 0 and 10. 0 μmol/L Cd Cl2 groups were significantly decreased than the 0. 0 μmol/L Cd Cl2 group( P < 0. 05). The proliferation rate in ICI group was lower than that of the non-ICI group( P < 0. 05). When Cd Cl2 concentration was 2. 5,5. 0 and 10. 0 μmol/L,the apoptosis rate of MCF-7 cells in non-ICI group increased compared with those cells without exposure to Cd Cl2( P < 0. 05). The relative expression of ERα,ERβ,miRNA-155 and miRNA-200 c increased( P < 0. 05). The proliferation of MCF-7 cells in ICI group decreased( P < 0. 05),and the relative apoptosis rate increased( P < 0. 05); and the relative expression of ERαand ERβ increased( P < 0. 05),the relative expression of miRNA-155 and miRNA-200 c decreased( P < 0. 05). When treated without Cd Cl2,the apoptosis rate of the ICI group increased compared with non-ICI group(P < 0. 05),the relative expression of ERα and ERβ decreased( P < 0. 05),and the relative expression of miRNA-155 and miRNA-200 c were increased( P < 0. 05). When Cd Cl2 concentration was 2. 5,5. 0 and 10. 0 μmol/L,the apoptosis rate and the relative expression of ERα,ERβ,miRNA-155 and miRNA-200 c decreased compared with the non-ICI group treated with same dose Cd Cl2(P < 0. 05). CONCLUSION: Cadmium can induce cell apoptosis and increase expression of miRNAs through the ER signaling pathway.

Effect of Anastrozole on Sex Hormone Levels and MCF-7 Cell Inhibition in Postmenopausal Women with Breast Cancer / 中国药师

Objective:To investigate the effect of anastrozole on sex hormone levels and michigan cancer foundation-7 ( MCF-7 ) cell inhibition in postmenopausal women with breast cancer. Methods: Totally 80 women with breast cancer were selected. Among them, 40 cases of patients received the routine management and tamoxifen treatment were included into the control group, and another 40 cases of patients received the routine management and anastrozole treatment were included into the observation group. The changes of sex hormone levels, effects and adverse drug reactions of two groups were compared between basetine and three months after treat-ment, meanwhile, the incidence of MCF-7 cell inhibition between two groups. Results:After treatment, the levels of estradiol( ER) , progesterone(PR) and luteotropic hormone(LH) were lower than baselime, however, the level of testosterone was contrary(P <0. 05). Meanwhile, the level of ER, PR and LH in control group were lower than baselime however, the level of testosterone was cont-ray (P<0. 05). The incidence of MCF-7 cell inhibillion between two grugs at time 72 h were higher than the 48 h, the incedence of MCF-7 cell inhibition in observation group were significantly higher than control groups at time 48 h and 72 h (P<0. 05). The rate of total effective in control group(45. 0%) was lower than the observation group(70. 0%)(P>0. 05). There was no significant differente in the micidence of adverse reactions. Conclusion:Anastrozole used for postmenopausal women with breast cancer has significant effi-cacy, high safety and promising inhibitory effect on MCF-7 cell. It can effectively adjust sex hormone level, reduce occurrence and transfer risks of breast cancer.

Inhibition effect of curcumin on proliferation and invasion of breast cancer cell by decreasing expression of MT1-MMP / 实用医学杂志

Objective To investigate whether MT1-MMP is involved in the inhibition effect of curcumin on the proliferation and invasion of breast cancer cell and the mechanism . Methods Firstly, MCF-7 cell lines transfected by MT1-MMP eukaryotic expression vector was established. We divided all cells into 3 groups,including null vector transfection group, non-transfected and transfected group with different concentrations of curcumin. The expression of MT1-MMP protein, the proliferation and invasion ability were respectively analyzed by western blot, transwell method, and cell counting kit-8 (CCK-8). Results The expression of MT1-MMP was inhibited by curcumin. Transwell and CCK-8 experiment indicated the proliferation and invasion abilities of MT1-MMP transfected MCF-7 cells were inhibited by curcumin in a concentration dependent manner. Conclusion The inhibition value of curcumin on proliferation and invasion is probably due to its ability to inhibit the expression of MT1-MMP.

Preparation of liposomal pemetrexed and its effects on breast cancer / 上海交通大学学报（医学版）

Objective · To prepare the liposomal pemetrexed and investigate its effects on MCF-7 breast cancer cells in vitro and nude mice bearing MCF-7 xenograft tumors. Methods · Liposomal pemetrexed was prepared by film dispersion method. Inhibition of MCF-7 breast cancer cell lines was evaluated by CCK-8 method, and anti-tumor effects were investigated on Balb/c nude mice bearing MCF-7 xenograft tumors. Results · Liposomal pemetrexed inhibited the growth of MCF-7 cells. When the concentrations of pemetrexed were 0.20, 0.40 and 10.00 μg/mL, the cell viability in experiment group (liposomal pemetrexed) was significantly lower than that in control group (pemetrexed of same concentration gradient), with P values of 0.013, 0.035 and 0.041, respectively. Compared with blank group (same volume of PBS), the volumes and weights of tumors of nude mice in experiment group(liposomal pemetrexed) and control group (same volume of pemetrexed) were significantly lower, and the volume and weight of tumor in experiment group were also significantly lower than those in control group (P=0.000). Conclusion · Compared to bulk drug of pemetrexed, liposomal pemetrexed can inhibit the growth of MCF-7 breast cancer cells and the Balb/c nude mice bearing MCF-7 xenograft tumors.

Selection and identification of oridonin high yield strain and its antitumor activity / 中草药

Objective: To select and identify the endophytic fungi of Rabdosia rubescens as well as study on its antitumor activity in vitro in order to obtain the high yield strains of oridonin. Methods: Using the methods of TLC and HPLC, the oridonin high yield strain was selected from 256 strains of endophytic fungi which were isolated from the previous experiments. Oridonin of fermentation broth was determined by HPLC-MS. The strain was identified by morphological characteristics and ITS sequence methods and its antitumor activity in vitro was detected by MTT assay. Results: An oridonin high yield strain M-J-5 was obtained and identified as Penicillium oxalicum, which can inhibit the activity of human breast cancer cell line MCF-7. Conclusion: a oridonin high yield strain is obtained with antitumor activity and provides scientific basis for microbial production of oridonin and the development of new anticancer drugs.