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Objective To analyze the regulatory of miRNA (miR)-149-5p for the expression of Aurora-B in esophageal squamous cell carcinoma (ESCC). Methods The pathologic histology paraffin blocks of 61 patients with ESCC in Shanxi Provincial People's Hospital from January 2010 to December 2017 were collected. The immunohistochemical staining and tissue section in situ hybridization method were used to observe the expressions of Aurora-B and miR-149-5p in the tumor tissues and adjacent mucosas of the patients, and their relationship with clinicopathological parameters was analyzed. miRNA was predicted by using software TargetScan and miR walk. The relationship between Aurora-B and miR-149-5p were verified by using Western blot in ESCC TE-1 cells. Results The result of immunohistochemical staining showed that in 61 patients, 37 (61％) tumor tissues showed higher expression of Aurora-B compared with adjacent mucosas, the Aurora-B expression in 15 (26％) tumor tissues were in line with benign tissues, the Aurora-B expression in 9 (13％) tumor tissues were inferior to benign tissues. The expression of Aurora-B were not correlated with age (χ2=0.008, P=0.929), gender (χ2=0.088, P=0.767), grade of differentiation (χ2=2.632, P=0.268), but correlated with TNM staging (χ2=15.153, P<0.01) and lymph node metastasis (χ2=5.979, P=0.014). The miR-149-5p was predicted to combine with Aurora-B 3'untraslated region (UTR) by using TargetScan and miRwalk software. The result of in situ hybridization showed that the miR-149-5p showed low expression in 22 (36％) tumor tissues. The expression of miR-149-5p was correlated with Aurora-B expression (χ2 = 5.622,P= 0.018), and not correlated with age (χ2= 2.617, P= 0.106), gender (χ2= 1.529, P= 0.216), grade of differentiation (χ2 = 2.854, P= 0.240), TNM staging (χ2 = 0.870, P= 0.351) and lymph node metastasis (χ2= 0.128, P= 0.351). The Western blot results of TE-1 cells showed that the expression of Aurora-B in simultaneous over-expression of miR-149-5p and Aurora-B group was higher than that in over-expression of miR-149-5p group, and lower than that in over-expression of Aurora-B group. Conclusion The miR-149-5p can be involved in ESCC progression through regulating the expressions of Aurora-B.
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Background and purpose:MicroRNA(miRNA) is a class of small non-coding RNA playing an important regulatory role in many tumors. This study investigated which miRNA might negatively regulate the expression of Aurora-B in osteosarcoma cells, and to lay the foundation for the further investigation of the effort and regulation of Aurora-B in osteosarcoma malignant phenotype.Methods:Bioinformatics prediction software (http://www.targetscan.org) and luciferase assays were used to investigate which miRNA might target to modulate the Aurora-B. Real-time fluorescent quantitative polymerase chain reaction (RTFQ-PCR) and Western blot assay were used to further verify which miRNA could negative regulate the expression ofAurora-B gene.Results:Bioinformatics prediction showed let-7 family have the possibility to modulate the expression of Aurora-B; Luciferase assays showed thatAurora-B might be the target gene of let-7a/b/c/d/e/f/g/i; RTFQ-PCR and Western blot analysis testiifed that both the expression levels of Aurora-B mRNA and Aurora-B protein were signiifcantly decreased in Let-7a/g/i up-regulated U2-OS and HOS cells, compared to the cells in the negative control group; but in Let-7b/c/d/e/f up-regulated U2-OS and HOS cells, the expression levels of Aurora-B mRNA and Aurora-B protein have no signiifcant difference, compared to the cells in the negative control group.Conclusion:Let-7a/g/i may downregulate the expression of Aurora-B in human osteosarcoma cells.
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Objective To study the effect of tetramethylpyrazine injection on proliferation and apoptosis of human acute lymphoblastic leukemia (ALL) cell line Jurkat and the relevant molecular mechanisms.Methods Cells were treated with tetramethylpyrazine injection at various concentrations (0,0.25,0.50,0.75,1.00,1.25 and 1.50 mg/ml),CCK-8 method was used to detect the inhibition rates at 24,48 and 72 h.After cells were treated with different concentrations of tetramethylpyrazine injection (0,0.50,1.00 and 1.50 mg/ml) for 48 h,the cell cycle and apoptosis were detected by flow cytometry,and the Aurora-B and Survivin protein expression were analyzed by Western blot.Results Compared with the control group (cells without tetramethylpyrazine injection treatment),various concentrations of tetramethylpyrazine injection could effectively inhibit Jurkat cells proliferation in a time-and dose-dependent manner (P ＜ 0.05),and IC50 at 24 h,48 h and 72 h after treatment were (1.33±0.16),(0.91±0.10) and (0.67±0.11) mg/ml,respectively.After cells were treated with tetramethylpyrazine injection at different concentrations (0.5,1.0 and 1.5 mg/ml) for 48 h,the number of treated cells in G2/M phase was increased,but that in S phase was decreased,and the apoptosis rates were significantly higher than that of control group,with a dose dependence (P ＜ 0.05).The results of Western blot showed that the expression levels of Aurora-B and Survivin in treated cells were lower than that of control group,also with a dose dependence (P ＜ 0.05).Conclusions Tetramethylpyrazine injection can effectively inhibit proliferation and induce apoptosis of Jurkat cells in vitro,and its underlying mechanisms involve with down-regulation of the Aurora-B and Survivin protein expression.
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Background and purpose: Aurora kinases, frequently detected to be over-expressed in some human tumors, regulate many essential events during tumor cell mitosis progression and have been regarded as potentially important targets for cancer therapy. This study was designed to detect Aurora-B expression in cervical carcinoma, explore the relation Aurora-B expression and clinicopathologic feature. So, Aurora-B kinase inhibition ZM447439 was used to investigate the effect of ZM447439 on SiHa cell line and the synergy effect with paclitaxel. Methods:Detected Aurora-B protein in cervical carcinoma(70 patients), CINⅡ(46 patients) and normal cervix(21patients) by immunohistochemistry. The inhibitory effects of ZM447439 and paclitaxel in SiHa cell line were investigated by MTT based assay, The expression of apoptosis associated protein was measured by Western blot. Results:The rate of Aurora-B expression is 71.43%in cervical cancer, with no signiifcant correlation to clinical stage, age, lymph node metastasis, vascular invasion (P>0.05). Aurora-A protein expression has significant correlation to pathological type and grade(P1.15). The level of P53 was increased significantly through ZM447439 treatments with Western blot. Conclusion: Our data provides strong evidence that high expression of Aurora-B in cervical cancer. The targeted Aurora-B inhibitors, ZM447439, can enhance paclitaxel anti-tumor activity in cervical cancer.
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Objective To observe the expression of survivin and Aurora B in human pancreatic cancer BXPC3 cells after the treatment of sulindac and to explore the potential mechanism. Methods MTr assay was used to determine the effect of sulindac on the proliferation of the BXPC3 cells. RT-PCR was used to detect the expression of mRNA level of survivin and Aurora B, western blot was used to detect protein expression of survivin and Aurora B Thr-232. Cell cycle and apoptosis were detected by flow eytometry (FCM). Results The BXPC3 cells were inhibited by sulindac in a dose and time-dependent manner; the expression of mRNA of survivin and Aurora B were both significantly decreased from 1.5644 and 0.6554 to 0. 4372 and 0.1132 (P< 0.01), the expression of survivin protein and the phosphorylation of Aurora B Thr-232 were also decreased from 1.2735 and 0.4680 to 0.2126 and 0.2546 (P<0.01); the proportion of cells in the G0/G1 phase was increased from (56.65±1.93)% to (70.58±3.21)% (P<0.01). Conclusions Sulindac had inhibitory effects on the growth of BXPC3 cells, the possible mechanism was via decreasing the expression of survivin which depressed the activity of Aurora B, then the CPC was influenced. The most of the cells were blocked in the G0/G1 phase, and the cells' mitosis was inhibited.
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The expression of Aurora B in normal endometria and endometrial carcinomas and its relation with clinicopathologic parameters of endometrial carcinomas were investigated. Streptavidin-biotin peroxidase (SP) immunohistochemical technique was used to detect the expression of Aurora B in 10 cases of normal proliferative phase endometria, 10 cases of normal secretory phase endometria and 72 cases of endometrial carcinomas respectively. According to the 1988 International Federation of Gynecology and Obstetrics (FIGO) grade, there were 37 patients in grade 1, 23 in grade 2 and 12 in grade 3 respectively. According to the FIGO stage, there were 59 patients in stage Ⅰ-Ⅱ and 13 patients in stage Ⅲ-Ⅳ. Aurora B was expressed in both normal proliferative phase endometria, secretory phase endometria and endometrial carcinomas, but its positive labeling index (PLI) in proliferative phase endometria was significantly higher than that in secretory phase endometria (P<0.01) and endometrial carcinomas (P<0.01). The PLI of Aurora B was lower in tumors with well differentiation (G1), low surgical staging (Ⅰ-Ⅱ), and≤1/2 myometrial invasion than that in tumors with moderate and low differentiation (G2-G3), higher surgical staging(Ⅲ-Ⅳ), and>1/2 myometrial invasion (all P<0.01). Aurora B exerts its functions in the replication of normal endometrial glandular cells; Expression of Aurora B is significantly correlated with biologic behavior of endometrial carcinoma, indicating that Aurora B may be a promising prognostic factor in endometrial carcinoma.