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Objective To investigate the correlation between the expression of the activator of HSP90 ATPase-1(AHA1),lysyl oxidase like-2 protein(LOXL2)in osteosarcoma tissues with mRNA expression of invasion and metastasis genes and their clinical significance.Methods A total of 90 osteosarcoma patients diagnosed and treated in North China Medical and Health Group Fengfeng General Hospital from February 2016 to March 2017 were selected as the research object.The expression of AHA1 mRNA,LOXL2 mRNA and invasion and metastasis genes Wnt family member 9A(Wnt9a)mRNA,zinc finger E-box binding homologous box 1(ZEB1)mRNA,zinc finger E-box binding homologous box 1(ZEB2)mRNA,N-cadherin(N-cad)mRNA,and vimentin(Vim)mRNA in tissues were detected by real-time fluorescence quantitative PCR.Pearson correlation analysis was used for correlation analysis.The differences in expression of AHA1 mRNA and LOXL2 mRNA in osteosarcoma patients among different clinical characteristics were compared.Kaplan-Meier survival analysis was used to analyze the effect of AHA1 mRNA and LOXL2 mRNA on the prognosis of osteosarcoma patients.The prognostic factors of osteosarcoma patients were analyzed by univariate and multivariate COX regression.Results The expressions of AHA1 mRNA(3.16±0.59),LOXL2 mRNA(2.84±0.44)and invasion and metastasis genes[Wnt9a mRNA(3.23±0.42),ZEB1 mRNA(2.73±0.39),ZEB2 mRNA(2.52±0.56),N-cad mRNA(2.71±0.65)and Vim mRNA(2.81±0.73)]in osteosarcoma tissues were higher than those in paracancerous tissues(1.10±0.21,0.95±0.18,0.79±0.15,0.64±0.11,0.98±0.19,0.68±0.14,0.72±0.15),and the differences were statissically significant(t=31.206,37.716,51.903,48.931,24.706,28.964,26.605,all P<0.05).There was a significant positive correlation between AHA1 mRNA and LOXL2 mRNA expression in osteosarcoma(r=0.712,P<0.05).The expressions of AHA1 mRNA and LOXL2 mRNA were significantly positively correlated with the expressions of invasion and metastasis genes(Wnt9a,ZEB1,ZEB2,N-cad,and Vim mRNA)in tumor tissue of osteosarcoma group(r=0.504～0.720,all P<0.05).The expressions of AHA1 mRNA and LOXL2 mRNA in osteosarcoma tissues with Eneeking stage Ⅲ,soft tissue infiltration,and lung metastasis were higher than those in patients with Eneeking stage Ⅰ～Ⅱ,no soft tissue infiltration,and no lung metastasis,with significant differences(t=14.122～171.054,all P<0.05).The 5-year survival rates of patients in the AHA1 mRNA high expression group and low expression group were 36.36%(16/44)and 78.26%(36/46),respectively.The 5-year cumulative survival rate of patients in the AHA1 mRNA high expression group was significantly lower than that in the low expression group(Log-rank χ2=16.081,P<0.05).The 5-year survival rates of patients with high and low expression of LOXL2 mRNA were 34.88%(15/43)and 78.72%(37/47),respectively.The 5-year cumulative survival rate of patients in the LOXL2 mRNA high expression group was significantly lower than that in the low expression group(Log-rank χ2=15.880,P<0.05).Lung metastasis(OR=1.921,P<0.05),Eneeking stage Ⅲ(OR=1.906,P<0.05),AHA1 mRNA high expression(OR=1.405,P<0.05),and LOXL2 mRNA high expression(OR=1.733,P<0.05)were independent risk factors affecting the poor survival prognosis of osteosarcoma patients.Conclusion The expressions of AHA1 mRNA and LOXL2 mRNA in osteosarcoma were increased,and they were correlated with the expression of invasion and metastasis genes,indicating they may be independent risk factors affecting the poor survival and prognosis of osteosarcoma patients.
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Objective@#Explore the function and regulatory mechanism of Annexin A1 (ANXA1) in bladder cancer cell proliferation, apoptosis and migration.@*Methods@#From February 2018 to June 2019, we use T24 cells as the model and divide it into over-expression control group (ctrl), ANXA1 over-expression group (ANXA1), knockdown control group (shctrl), ANXA1 knockdown group 1 (shANXA1-1), ANXA1 knockdown group 2 (shANXA1-2) and ANXA1 knockdown group 3 (shANXA1-3). 24 hours after the culture, the cells were collected and the mRNA expression level of ANXA1 was detected by Real-Time quantitative PCR. The cell activity was detected by CCK-8; the cell apoptosis and cycle were detected by flow cytometry. The cell migration was detected by Transwell assay.@*Results@#The Real-Time quantitative PCR showed that the expression of ANXA1 in the over expression group was significantly higher than that in the over expression control group (15 369.00±874.20 and 1.00±0.07, P<0.001). The expression of ANXA1 in the knockdown group 2 and 3 were significantly lower than that in the knockdown control group (0.51±0.04, 0.51±0.02 and 1.00±0.04, P<0.001). Compared with the over expression control group(1.61±0.01), the cell activity of the over expression group(2.04±0.02)was significantly increased (P<0.001), while the activity of the knockdown group 2 and 3 (1.40±0.002 and 1.31±0.003)were significantly decreased than the knockdown ctrl group (1.73±0.01)(P<0.001). The results of flow cytometry showed that the number of G0/G1 cells in the over-expression group was significantly lower than that in the over-expression control group (28.14±0.33 and 46.19±0.73, P<0.001), while that in the knockdown group 2 and 3 were significantly higher than that in the knockdown control group (58.670±0.49, 62.34±4.01 and 45.59± 0.19, P<0.001 and P<0.05). There was no significant difference in the number of apoptosis between the over-expression group and the over-expression control group (P>0.05), while the number of apoptosis in the knockdown group 2 and 3 were significantly higher than that in the knockdown control group (13.04%, 14.58% and 7.76%, P<0.001). Cell function analysis showed that the number of cells passing through the membrane of the over expression group was significantly higher than that of the over expression group (525.00±9.30 and 385.70±13.40, P<0.01), while that of the knockdown group 2 and 3 were significantly lower than that of the knockdown control group (214.70±6.40, 226.00±5.30 and 398.70±10.00, P<0.001).@*Conclusions@#Over-expression of ANXA1 significantly promoted the proliferation, cycle and migration of T24 cells and inhibited apoptosis. On the contrary, ANXA1 knockdown inhibited the proliferation, cycle and migration of T24 cells and promoted apoptosis.
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Objective Explore the function and regulatory mechanism of Annexin A1 (ANXA1) in bladder cancer cell proliferation,apoptosis and migration.Methods From February 2018 to June 2019,we use T24 cells as the model and divide it into over-expression control group (ctrl),ANXA1 over-expression group (ANXA1),knockdown control group (shctrl),ANXA1 knockdown group 1 (shANXA1-1),ANXA1 knockdown group 2 (shANXA1-2) and ANXA1 knockdown group 3 (shANXA1-3).24 hours after the culture,the cells were collected and the mRNA expression level of ANXA1 was detected by Real-Time quantitative PCR.The cell activity was detected by CCK-8;the cell apoptosis and cycle were detected by flow cytometry.The cell migration was detected by Transwell assay.Results The Real-Time quantitative PCR showed that the expression of ANXA1 in the over expression group was significantly higher than that in the over expression control group (15 369.00 ± 874.20 and 1.00 ± 0.07,P ＜ 0.001).The expression of ANXA1 in the knockdown group 2 and 3 were significantly lower than that in the knockdown control group (0.51 ± 0.04,0.51 ± 0.02 and 1.00 ± 0.04,P ＜ 0.001).Compared with the over expression control group (1.61 ± 0.01),the cell activity of the over expression group(2.04 ± 0.02) was significantly increased (P ＜ 0.001),while the activity of the knockdown group 2 and 3 (1.40 ± 0.002 and 1.31 ± 0.003) were significantly decreased than the knockdown ctrl group (1.73 ± 0.01) (P ＜ 0.001).The results of flow cytometry showed that the number of G0/G1 cells in the over-expression group was significantly lower than that in the over-expression control group (28.14 ± 0.33 and 46.19 ± 0.73,P ＜ 0.001),while that in the knockdown group 2 and 3 were significantly higher than that in the knockdown control group (58.670 ± 0.49,62.34 ± 4.01 and 45.59 ± 0.19,P ＜ 0.001 and P ＜ 0.05).There was no significant difference in the number of apoptosis between the over-expression group and the over-expression control group (P ＞ 0.05),while the number of apoptosis in the knockdown group 2 and 3 were significantly higher than that in the knockdown control group (13.04％,14.58％ and 7.76％,P ＜ 0.001).Cell function analysis showed that the number of cells passing through the membrane of the over expression group was significantly higher than that of the over expression group (525.00 ± 9.30 and 385.70 ± 13.40,P ＜ 0.01),while that of the knockdown group 2 and 3 were significantly lower than that of the knockdown control group (214.70 ± 6.40,226.00 ± 5.30 and 398.70 ± 10.00,P ＜ 0.001).Conclusions Over-expression of ANXA1 significantly promoted the proliferation,cycle and migration of T24 cells and inhibited apoptosis.On the contrary,ANXA1 knockdown inhibited the proliferation,cycle and migration of T24 cells and promoted apoptosis.
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Objective:To investigate the expression of Slug,EMMPRIN and E-cadherin in salivary adenoid cystic carcinoma (SACC)and its correlation with clinicopathological characteristics,and the correlation among themselves.Methods:Slug,EMMPRIN and E-cadherin expression in 1 1 5 SACC cases of SACC was examined by immunohistochemical staining.The results and clinicopatho-logical data were statistically analyzed.Results:High positive expression frequencies of Slug(76.5%)and EMMPRIN(69.6%)and low positive expression frequency of E-cadherin(51 .3%)were found in 1 1 5 SACC cases.The expression of Slug and EMMPRIN was positively associated with the histopathological types,clinical stages,perineural invasion,recurrence and distance metastasis(P <0.05).The expression of E-cadherin was negatively associated with the histopathological types,clinical stages,perineural invasion and distance metastasis(P <0.05).There was a significant correlation between Slug and EMMPRIN expression(P <0.05),negative correlation between EMMPRIN and E-cadherin expression(P <0.05)and between Slug and E-cadherin expression(P <0.05).Con-clusion:The expression of Slug,EMMPRIN and E-cadherin is closely correlated to the clinicopathological characteristics of SACC.