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Objective To investigate the role of estrogen and progesterone in the tumorigenesis of triple-negative breast cancer and its molecular mechanism. Methods To construct of MDA-MB-231/miR-145 and MDA-MB-231/ miR-SCR(scramble control) stable cell lines. The cells were cultured in five groups that were respectively miR-SCR group, miR-145 group, miR-145 + E2 group, miR-145 + P4 group, miR-145 +E2 +P4 group, grouped adminis-tradion. Cell proliferation was detected by CCK-8 kit, the expression level of miR-145 target gene insulin-like growth factor receptor-1 (IGF-IR), N-RAS was detected by Western blot, expression of IGF-1R, N-RAS downstream signaling molecule vascular endothelial growth factor(VEGF) and expression of miR-145 after transfection was detected by real-time quantitative PCR(qRT-PCR). Results The results of CCK-8 assay showed that estrogen and progesterone combined administration could reverse the inhibitory effect of miR-145 on the proliferation of triple-negative breast cancer cells (P ＜ 0. 05 ). The results of Western blot showed that estrogen and progesterone combined administration could increase the expression level of IGF-1 R and N-RAS in the target gene of miR-145. The results of qRT-PCR showed that the expression level of miR-145 was significantly higher than that of control group (P ＜ 0. 05) and the expression of VEGF was significantly increased by estrogen and progesterone combined administration (P ＜0. 01). Conclusion Estrogen and progesterone combined administration can promote the expression of VEGF and cell proliferation, and then reverse the anti-tumor effect of miR-145 in the triple-negative breast cancer.
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Objective To explore the mechanism of notch signaling pathway in the treatment of cervical cancer with notch receptor blocker(DAPT).Methods After treatment of Me180 cells with different concentrations of DAPT combined with curcumin-photodynamic therapy (PDT),3-(4,5 Dimethylthiazlo-yl)-2,5 Dimethylthiazol-2-yl)-2,5 dipheny tetrazo lium bromide(MTT)assay was used to detect the effect on the proliferation of Me180 cells,annexin V-FITC/PI combined with flow cytometry was used to detect the effect on apoptosis,and real-time fluorescence quantitative PCR and Western blot were used to detect the mRNA and protein expressions of Notch1 gene respectively.Results Both DAPT(1.0 μmol/L)and curcumin-PDT groups could inhibit the proliferation and induce apoptosis of cervical cancer cell line Me180,and the synergistic effect of the two groups was significant (P＜0.05 or＜0.01).DAPT and curcumin-PDT group reduced the mRNA expression level of Notch1 in cervical cancer Me180 cells with inhibition ratio of 40.0％ and 32.3％ respectively.DAPT and curcumin-PDT group inhibited the protein expressions of Notch1,nuclear factor-kappa B(NF-kB),vascular endothelial growth factor (VEGF),and the synergistic effect of the two groups was statistically significant.Conclusions Both DAPT and curcumin-PDT may effectively block the conduction of Notch signaling pathway,which is associated with down-regulation of the expression of Notch1 and NF-kappa B.Notch signaling pathway may be one of the targets of curcumin photodynamic therapy.
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binding,cellular organelle membrane,and cellular metabolic process of GO enrichment.For the KEGG pathways, the target genes mainly concentrated on the focal adhesion pathway.Conclusion There is a different expression of novel microRNA between SLE and NC groups.The target genes from differently-expressed novel microRNA may play an important role in the pathogenesis of SLE and clinical symptoms and may be the unique target for further research.
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Objective To study the effects of curcumin mediated photodynamic therapy (PDT)on the growth and proliferation in human cervical cancer cell line H8 in vitro and in vivo,and to investigate its antitumor mechanisms.Methods The effects of curcumin mediated PDT on proliferation of human cervical cancer H8 cell by MTT assay was used to screen the optimal parameter.Changes in cell morphology were observed by May-Gr ünwald-Farbstoff Giemsa staining.The apoptosis rate was estimated by flow cytometry.The effect of PDT by curcumin on the expressions of Bcl-2,P53 and survivin in H8 cells was detected by fluorescence real-time reverse transcription-polymerase chain reaction (RT-PCR).Forty BALB/C nude mice underwent subcutaneous injection of H8 cell line so as to establish animal models,and then were randomly divided into four equal groups:control group,irradiation alone group,curcumin alone group,curcumin PDT group.HE staining and pathological examination were performed.Immunohistochemical study was conducted to detect the protein expression of the apoptosis inhibiting genes of Bcl-2.Results The proliferation inhibition of H8 cells was obvious after PDT when curcumin 5μmol/L with irradiation 100 J/cm2,and with dose dependent manner.Typical morphologic features of apoptosis appeared characterizedly by marked chromatin condensation,nuclear pyknosis and fragmentation,and the appearance of apoptotic bodies.The total apoptosis rate was higher in PDT group [(47.21 ± 4.11)％]than in control group(1.71 ±0.16) ％ (P＜0.01).The mRNA expression of Bcl-2,P53 and survivin in H8 cells were suppressed significantly.HE staining showed remarkable subcutaneous necrosis in the PDT group.Immunohistochemistry showed remarkable down-regulation of protein expression of Bcl-2(P＜0.01).Conclusions Curcumin-mediated photodynamic therapy has a significant killing effect on H8 cells in vivo and in vitro.Its antitumor effect might be related to induction of Tumor cell apoptosis and suppression of Bcl-2 mRNA and protein expression.