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Objective:To explore the effect of the mucin modulator Talniflumate (Tal) on breast cancer cells and its synergistic effect after combined with the chemotherapy drug paclitaxel (PTX).Methods:The breast cancer cells were cultured in vitro. Lymphocyte proliferation activity assay (MTS) was used to detect the effects of different concentrations of Talniflumate alone and paclitaxel on the survival rate of breast cancer cells. The effects of the above drugs on the apoptosis of breast cancer cells were detected by flow cytometry. Western blot was used to detect the expression of glucosamine transferase (GCNT3) (the target of Talniflumate) in breast cancer cells before and after the treatment with Talniflumate. Transcriptome sequencing clarified the changes in related signaling pathways after treatment with Talniflumate. Results:Talniflumate promoted the apoptosis of breast cancer cells MCF7 and MDA-MB-231 cells in a concentration-dependent manner. The combination of Talniflumate and paclitaxel had a significant synergistic killing effect in MCF7 cells but not MDA-MB-231 cells. Western blot indicated that GCNT3 was highly expressed in MCF7 cells, while almost no expression in MDA-MB-231 cells; Talniflumate could reduce the expression of GCNT3 in MCF7 cells and after combined with paclitaxel, the expression of GCNT3 was downregulated more significantly. Transcriptome sequencing suggested that Talniflumate can regulate the expression of multiple signaling pathways such as TNF, p53, and SNARE.Conclusions:Talniflumate could induce apoptosis of breast cancer cell. Talniflumate combined with paclitaxel has a significant synergistic effect in killing tumor cells in breast cancer cells with high GCNT3 expression such as MCF7. The mechanism of Talniflumate induce apoptosis of breast cancer cells may be related to multiple signaling pathways such as TNF, p53, and SNARE.
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Objective:APOBEC3B (A3B) is an important member of the apolipoprotein B mRNA editing enzyme catalytic polypeptide (APOBEC) family. The study aims to investigate the relationship between A3B expression and prognosis as well as resistance to cisplatin in non-small cell lung cancer (NSCLC).Methods:Real-time quantitative polymerase chain reaction (qRT-PCR) was used to analysis the A3B mRNA expression in 40 NSCLC tissues; Kaplan Meier plotter was used to analyse the correlation between A3B expression and clinical prognosis; in addition, the knock-down A3B expression cell line in human lung adenocarcinoma cell A549 was constructed; MTS and plate cloning experiment were performed to observe the changes in cell cisplatin sensitivity, and γ-H2AX immunofluorescence was used to quantitate the DNA damage.Results:Compared with adjacent tissues, A3B was highly expressed in NSCLC tissues (27/40). Kaplan Meier plotter analysis showed that A3B expression was positively correlated with NSCLC overall survival (OS) [adenocarcinoma: HR=0.64(0.47-0.86), P=0.002 6; squamous cell carcinoma: HR=0.77(0.59-1.01), P=0.006]. Cell-based studies showed that the knock-down A3B expression contributed to sensitivity to cisplatin in A549 cells. Conclusions:A3B mediates the sensitivity of lung cancer to cisplatin. This effect may partly explain why NSCLC patients with high A3B expression have a better prognosis.
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Objective APOBEC3B (A3B) is an important member of the apolipoprotein B mRNA editing enzyme catalytic polypeptide (APOBEC) family.This study aimed to investigate its important role in the metastasis of small cell lung cancer (NSCLC).Methods The statistical relationship between A3 B and clinicopathological data was analyzed in 249 cases of NSCLC.Sanger sequencing was used to detect mutations in exon 5,6,7 and 8 of P53 in 74 cases of lung cancer.A3B overexpression cell line was constructed in human lung adenocarcinoma cells HCC827 to observe the change of cell migration and metastasis capacity.Results A3B was highly expressed in NSCLC tissues compared with normal lung tissues.The expression of A3B was closely related to the lymph node metastasis of NSCLC and the mutation rate of p53 was positively correlated with the expression of A3B.In vitro experiment,it showed enhanced migration and increased metastatic potential in cells after overexpression of A3B.Conclusions A3B-mediated mutations in P53 may play a key role in the metastasis of NSCLC.
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Objective To study the role of leucine rich repeat neuronal 3 (LRRN3) in the prolif-eration of non-small cell lung cancer and its possible mechanism of expression regulation. Methods The expression of LRRN3 in non-small cell lung cancer was detected by real-time quantitative polymerase chain reaction ( qPCR) , immunohistochemistry and bioinformatics retrieval;A lung cancer cell line A549-LRRN3 with stable over-expression of LRRN3 was established by lentivirus over-expression technology;The effect of LRRN3 on the proliferation of non-small cell lung cancer was detected by methyl thiazolyl tetrazolium ( MTT) assay;Bioinformatics search for changes in methylation of LRRN3 promoter region and treatment of lung cancer cells by methyltransferase inhibitors to detect the effect of methylation on the regulation of LR-RN3 expression; Finally, bioinformatics search analyzes the correlation between LRRN3 and lung cancer prognosis. Results The mRNA expression of LRRN3 in clinical tissues of non-small cell lung cancer (n=12) was significantly lower than that of adjacent normal tissues (n=12) (P=0. 0014). The results of immunohistochemistry showed that the protein expression level of LRRN3 in non-small cell lung adenocar-cinoma was lower than that in normal tissues (P=0. 001), and the expression in non-small cell lung squa-mous cell carcinoma was also lower than that in normal tissues (P=0. 003). Overexpression of LRRN3 in-hibited the proliferation of tumor cells (P<0. 01), and the hypermethylation of LRRN3 in the promoter re-gion inhibited its transcriptional expression. LRRN3 was positively correlated with the survival prognosis of lung adenocarcinoma (P=5. 2e-09;HR=0. 48). Conclusions Hypermethylation in the promoter region of LRRN3 inhibits its transcriptional expression, thereby promoting the proliferation of lung cancer cells.
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Objective To investigate the effect of histone deacetylase inhibitor on Her2 positive breast cancer cell line BT474 and SKBR3 in apoptosis and metastasis.Methods Histone deacetylase inhibitor MS-275,suberoylanilide hydroxamic acid (SAHA)(4 μmol/L,and 50 μmol/iL,respectively) treated the cell lines BT474 and SKBR3 cells.Flow cytometer examined the apoptosis ratio.Transwell tested their metastatic activity.Western blot assay was performed to detect the associated proteins.Results SAHA and MS-275 inhibited the cell survival.The BT474 cell survival was (39 ± 11) ％,(54 ± 8) ％,and the SKBR3 survival was (62 ± 6) ％,(71 ± 9) ％,according to the fluorescence-activated cell sorting (FACS) result.SAHA and MS-275 induced the BT474 cell apoptosis 8.46± 0.28 (P ＜0.01),4.15 ± 0.71 (P ＜0.01) fold change,respectively;and upregulated the SKBR3 cell apoptosis ratio 5.51 ± 1.24 (P ＜0.01),4.04 ±0.69 (P ＜0.01) fold.The Transwell result showed that SAHA,MS-275 inhibited the Transwell ability of BT474 from the control 184.7 ± 18.8 to 104.3 ± 7.1,131.3 ±9.1 per view,and the SKBR3 from control 60 ± 16.7 per view to 14.3 ± 6.5,34.3 ± 8.7 per view.The Western blot result showed that SAHA,MS-275 inhibited the protein level of vimentin,Her2,β-catenin,histone deacetylase inhibitor (HDACi),and upregulated the acetylation level of histone 3.The E-caherin protein was regulated in BT474 and SKBR3 cells.Conclusions MS-275,SAHA can induce BT474 and SKBR3 apoptosis significantly,also inhibit their metastatic activity.
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] Objective To explore the role and mechanisms of FGF2 in chemo?resistance in breast cancer. Methods The inhibitors for different signal pathway were used to treat two drug?resistant breast cancer cell lines MCF?7/5?Fu and T47D/5?Fu established in our lab. MTS assay was used to determine chemo?sensitivity and Hoechst stain was used to measure apoptosis. Protein activation and FGF2 protein level in cell culture medium were detected by western blot and ELISA respectively. Results Akt inhibitor MK?2206 (20 nM) and mTOR inhibitor AZD8055 (2 nM) significantly reversed the chemo?resistance of MCF?7/5?Fu and T47D/5?Fu cell lines to 5?Fu and paclitaxel, but ERK1/2 inhibitor SCH772984 showed no significant effect. Compared to parent cell lines MCF?7 and T47D, p?Akt and p?S6K (represented as mTORactivity) levels were obviously up?regulated in MCF?7/5?Fu and T47D/5?Fu cell lines, and so do the FGF2 mRNA level and FGF2 protein level from culture medium. Moreover, FGFR inhibitor AZD4547 (4 nM) markedly reversed the chemo?resistance of MCF?7/5?Fu and T47D/5?Fu cell lines to 5?Fu and paclitaxel and down?regulated activation of FGFR?Akt?mTOR signal pathway. In agreement, FGF2 protein (10ng/ml) enhanced the chemo?resistance of MCF?7 and T47D cell lines to 5?Fu and paclitaxel and up?regulated activation of FGFR?Akt?mTOR signal pathway. Conclusion Activation of FGF2?FGFR?Akt?mTOR signal pathway promoted chemo?resistance of breast cancer cells.
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Objective To explore the role of xCT in the metastasis and drug resistance of nasopharyngeal carcinoma (NPC),to provide new evidences for the mechanism of NPC metastasis and drug resistance,and to enhance effects of chemotherapy.Methods Fluorescence quantitative polymerase chain reaction (PCR) and Western blot were used to detect the xCT expression of 5-8F and 6-10B.The xCT eukaryotic expression vector was constructed to transient transfect 6-10B cell lines.The 5-8F cell lines were treated with xCT antisense oligodeoxynucleotide (ASO).Scratching assay and transwell method were used to detect the cell metastasis and invasion abilities.The expressions of matrix metalloproteinase 1 (MMP1) of all the cell lines were surveyed by Western blot.Results The results of fluorescence quantitative PCR and Western blot showed that the expression of xCT in 5-8F was higher than 6-10B in the levels of mRNA and protein.Exogenous overexpression of xCT enhanced metastasis and invasion abilities of 6-10B cells.Silencing and inhibition of xCT could decrease metastasis and invasion abilities of 5-8F cells.The expressions of MMP1 protein in 5-8F were higher than 6-10B,and they were positively correlated with the expression of xCT.Conclusions xCT is closely related to the metastasis and invasion abilities of nasopharyngeal carcinoma.xCT could enhance the metastasis and invasion abilities of NPC cells.xCT might mediate proliferation and matastasis/invasion of NPC cells through MMP1.
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Objective To investigate the expression of hypoxia inducible factor 1?(HIF-1?),vascular endothelial growth factor(VEGF) and micro vessel density(MVD) in gastric carcinoma and to explore their correlation with clinical pathological features such as cancer invasion and metastasis.Methods Forty-eight samples of gastric carcinoma tissues were examined for the expression of HIF-1?,VEGF and CD34 by immunohistochemical method.Results The positive expression rates of HIF-1? and VEGF were 66.7% and 60.4% in gastric carcinoma respectively.The mean value of MVD was 42.5?14.7 in gastric carcinoma.The expressions of HIF-1?,VEGF and the value of MVD were significantly correlated with the depth of invasion,lymph node metastasis and TNM stage.The HIF-1? expression was positively correlated with VEGF expression and MVD value.Conclusion The overexpression of HIF-1?,VEGF and MVD consist in gastric carcinoma tissue.The HIF-1? expression is positively correlated with VEGF expression and MVD value.The overexpression of HIF-1?,VEGF and MVD value are closely related with invasion,metastasis and poor biological behavior of gastric carcinoma.