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Objective To investigate the effects of chemotherapeutic drugs on ER-α expression and methylation in breast cancer cells.Methods Human breast cancer cells MCF-7(ER+,Luminal A) were induced by paclitaxel(PTX) and epirubicin(EPI) for more than 6 months,with an incremental dose,respectively.The expression and methylation status of ER-α in MCF-7 cells were detected before and after drug treatment.miRNAs with consistent expression changes in MCF-7 cells after two drugs' treatment were screened by microarray,and verified by quantitative PCR (qPCR).Targets of the most significantly down-regulated miRNA were analyzed by bioinformatics.miRNA inhibitor was transfected into MCF-7 cells,miRNA mimic was transfected into MCF-7/PTX and MCF-7/EPI cells,then ER-α and DNA methyltransferase 1 (DNMT1) expression were detected by Western blot,and ER-α methylation was detected by quantitative methylation-specific PCR (qMSP).Results PTX resistant MCF-7/PTX cell line and EPI resistant MCF-7/EPI cell line were established.Both drug treatments caused a decrease in ER-α protein expression and an increase in methylation levels,with up-regulation of DNMT1 and his tone deacetylase 1 (HDAC 1) expression.miRNAs with consistent expression changes in MCF-7 cells after drug treatments were screened and verified by qPCR,the most significant down-regulation among which was miR-148b.Bioinformatics analysis,and further confirmed by luciferase reporter gene assay (Luciferas) that DNMT1 was a direct target of miR-148b.miR-148b inhibitor induced decreased expression of ER-α and increased methylation level in MCF-7 cells,accompanied by increased expression of DNMT1;whereas miR-148b mimic caused an increased expression of ER-α and decreased methylation level in MCF-7/PTX and MCF-7/EPI cells,with a decreased expression of DNMT1.Conclusion Chemotherapeutic drugs (represented by PTX and EPI) induce aberrant miRNA expression in breast cancer MCF-7 cells,and down-regulate miR-148b further to attenuate the inhibition of DNMT1 expression,which promote,hypermethylation and down-regulation of ER-α.
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Objective:To investigate the pharmacodynamics and mechanism of Chaijin Sanjie prescription (CJSJP) on rat mammary gland hyperplasia, in order to provide experimental basis for the research and development of new Chinese medicine. Method:SD rat model of mammary gland hyperplasia was established through exogenous intramuscular injection with estrogen and progesterone. After successful establishment of the model, the rats were randomly divided into normal group, model group, and low, medium and high-dose CJSJP groups (3.13, 6.26, 12.52 g·kg-1) and Rupixiao (0.517 g·kg-1) group, with 9 rats in each group. After 28 days of administration, estradiol (E2), progesterone (P) and rolactin (PRL) were measured by radioimmunoassay, uterus and ovary coefficients were calculated; nipple diameter and breast histopathology were observed, estrogen receptor-α(ER-α) expression in mammary gland was measured by immunohistochemistry, and gonadotropin-releasing hormone (GnRH) and gonadotropin-releasing hormone receptor (GnRH-R) mRNA expressions in hypothalamus, pituitary were measured by Real-time PCR. ICR mice were randomly divided into normal group, low, medium and high-dose CJSJP groups (5.2,10.4,20.8 g·kg-1) and Luotongding group (0.038 6 g·kg-1) according to their body weight. Twelve mice in each group were given drugs for 7 days, and 0.6% acetic acid was injected intraperitoneally for 30 minutes after the last administration. The writhing times were observed within 15 minutes. Result:Compared with the normal group, the diameter of nipple was widened, serum E2 was significantly increased (Pα expression were increased in model group. compared with model group, the diameter of nipple was significantly decreased in high dose group of CJSJP (P2 was decreased in all dose groups of CJSJP, pathological score of breast hyperplasia was decreased in middle and high dose groups of CJSJP, GnRH mRNA in hypothalamus was decreased in all dose groups of CJSJP. The writhing times of mice in high dose group of CJSJP was decreased (PConclusion:Chaijin Sanjie prescription can improve the lesions of breast hyperplasia. The therapeutic mechanism may be related to the regulation of GnRH gene expression in hypothalamus and the decrease of estrogen receptor expression.
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Objective To investigate the effects of ER-α36 on invasion of human gastric cancer cell lines SGC7901 by miR-143. Methods Lentiviral vectors were constructed to generate up-and down-regulations of miR-143 lentiviruses (LV-miR-143 and LV-anti-miR-143,respectively).The viruses were used to infect human gastric cancer cell lines SGC7901.The ER-α36 protein expression level and the invasion of constructed cells were detected by Western blot and transwell. The target gene of miR-143 was predicted by bioinformatics tools.Luciferase reporter assay was carried out to confirm the predicted target gene. Results The infection efficiency of the lentivirus titers of LV-miR-143 and LV-anti-miR-143 were over 80% shown by the green fluorescence. The ER-α36 expression level,the cell invasion in LV-miR-21 group were significantly lower than those in LV-anti-miR-21 group(P<0.05). Conclusions miR-143 plays an important role in the negative control of gastric cancer invasion by the regulation of ER-α36.
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Objective The aim of this study was to explore the correlation between postoperative aromatase inhibitor(AIs) -based bone metabolism and ERα gene rs9340799,rs2234693 single nucleotide polymorphisms(SNPs)in breast cancer.Methods One hundred and sixty-six breast cancer patients who underwent AIs treatment(≤2 years)were enrolled and hospitalized in our hospital from October 2015 to April 2017.The ERα gene rs9340799 and rs2234693 sites were sequenced and compared subtype of lumbar spine and femur,bone mineral density BMD value and the relationship between BMD value and T value.Results The BMD of lumbar spine in patients with ERα gene rs9340799 was significantly different when compared to those of A/A,A/G and G/G(P<0.01).The BMD of lumbar spine in patients with A/A and A/G genotypes were significantly higher than those in G/G genotypes(P<0.05).The BMD of lumbar spine in patients with ERα gene rs2234693 was significantly different when compared to those of T/T,T/C and C/C(P<0.01). The BMD of lumbar spine in patients with T/T and C/T genotypes were significantly higher than those in C/C genotypes(P<0.05). However,there was no difference in femoral BMD,lumbar spine,and femur T between the 2 subtypes of patients with genotypes(P>0.05).Conclusion Aromatase inhibitor-related bone loss(AIBL)may be related to ERα gene phenotype.In ERα gene rs9340799 and rs2234693 loci,C and G alleles may be susceptible genes for aromatase inhibitor-related bone loss(AIBL).
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Estrogen receptor-alpha 36 (ER-α36), which is a new type of ER with a relative molecular weight of 3.57×104, has been demonstrated to have many important physiological functions in different tissues and organs in recent years. Many studies have shown that the occurrence and development of cancer are closely related to the expression level of ER-α36, which suggests that ER-α36 may serve as a potential therapeutic target in the treatment of some cancers. This paper reviews the advances in research on the correlation between ER-α36 and some cancers such as breast cancer, gastric cancer, liver cancer, and so on.
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Purpose To detect the expressions of stress induced phosphoprotein 1 (STIP1) and estrogen receptor-α(ER-α) in papil-lary thyroid carcinoma and to analyse the relationship between STIP1 and ER-α. Methods 54 cases of paraffin-embedded tissues of papillary thyroid carcinoma, 18 cases of papillary thyroid carcinoma with lymph node metastasis, 15 cases of Hashimoto’ s thyroiditis, 10 cases of adjacent normal thyroid tissue were collected. The expressions of STIP1 and ER-αwere detected by immunohistochemistry, and the relationship between the expressions and clinicopathological features of papillary thyroid carcinoma was analyzed. Results The expression of STIP1 and ER-α in papillary thyroid cancer group ( 55. 6% and 44. 4%) were higher than that of normal thyroid group (10% and 0) and Hashimoto’s thyroiditis group (8. 3% and 0, all P0. 05). The expressions of STIP1 and ER-α in papillary thyroid carcinoma were not related to patients’ age , tumor location, number of tumors, tumer size, invasion of capsule, the concomitant Hashimoto’ s thyroiditis and TPO-Ab ( all P>0. 05). And the expressions of STIP1 was not related to gender, TG-Ab and the merger of nodular goiter (all P>0. 05). A positive correlation was found between the expressions of STIP1 and ER-αin thyroid papillary carcinoma (P<0. 05). Conclusion STIP1 and ER-α in papillary thyroid carcinoma may be related with lymph node metastasis.
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Objective To explore the effect of total flavonoids of Epimedium sagittatum (TFE) on the mRNA expressions of the estrogen receptor alpha (ERα) and estrogen receptor beta(ERβ) in hippocampus and hypothalamus in ovariectomized (OVX) Sprague-Dawley(SD) rats, and the mechanism of TFE against postmenopausal osteoporosis. Methods Forty-eight female SD rats,aged 10-11 months old, were randomly divided into 4 groups: a sham group, an ovariectomy group (rats were bilaterally ovariectomized), a TFE group, and a 17β-estradiol group (rats were fed with TFE and 17β-estradiol for 4 months, respectively). The RT-PCR was performed to determine the mRNA expression of ERα and ERβ in hypothalamus and hippocampus. Results Serum estradiol level, bone mineral density (BMD) of vertebra,wet weight of uterus, and the mRNA expressions of ERα and ERβ in hypothalamus and hippocampus were markedly decreased in OVX rats, all of which were reversed by 17β-estradiol treatment except the mRNA expression of ERβ. Similar results were achieved by TFE treatment except the wet weight of uterus. Conclusion TFE can improve BMD of vertebra in the OVX rats without side effects on the uterus. The mechanism may be related to increasing the mRNA expressions of ERα and ERβ in hypothalamus and hippocampus.