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Objective To explore the role of microtubule actin cross-linking factor 1 (MACF1) in the metastasis of gastric cancer.Methods From 2009 to 2012,at The First Affiliated Hospital of Zhengzhou University,the paraffin blocks of gastric cancer and normal tissue adjacent to cancer of 107 patients who received radical gastrectomy were collected.The expression of MACF1 in tissues at protein level was detected by immunohistochemical staining.In 2017,at The First Affiliated Hospital of Zhengzhou University,fresh specimens samples of gastric cancer and normal tissue adjacent to cancer of 42 patients who received radical gastrectomy were also collected.The expression of MACF1 at mRNA level was determined by quantitative real-time polymerase chain reaction (PCR).MACF1 knockout gastric cell line was established.The effects of MACF1 on cell migration and invasion were verified by wound-healing test and Transwell assay.The effects of MACF1 on cell microtubule and actin were analyzed by filamentous actin (F-actin) staining.T-test,chi-square test and multivariate analysis were used for statistical analysis.Results The positive expression rate of MACF1 in gastric carcinoma tissues was 71.0％ (76/107),which was significantly higher than that of the corresponding normal tissues adjacent to cancer (22.4％,24/107),and the difference was significant (t =4.145,P =0.016).The expression of MACF1 at mRNA level in cancer tissues of 42 patients with gastric cancer was 6.463 ±0.672,which was significantly higher than that of corresponding normal tissue adjacent to cancer (1.727 ± 0.331),and the difference was statistically significant (t =6.326,P ＜ 0.01).The differences in positive expression rate of MACF1 in different tumor infiltration depth,different TNM stage and lymph nodes metastasis were statistically significant (x2 =1.170,7.959 and 5.288;all P ＜ 0.01).The five-year survival rate of patients with high expression of MACF1 was 32.9％ (25/76),which was significantly lower than that ofpatients with normal MACF1 expression (64.5％,20/31),and the difference was statistically significant (x2 =25.093,P =0.034).The high expression of MACF1 was an independent prognostic factor affecting overall survival rate in patients with gastric cancer after surgery (hazard ratio (HR) =0.513,95％ confidence interval (CI):0.411 to 0.922,P =0.038).The results of wound-healing assay showed that at 24 hour after wound the migration ability of MACF1 knockout AGS-MACF1 / cells was (18.77 ± 3.82) ％,which was lower than that of wild type AGS cells ((76.24 ± 5.36) ％),and the difference was statistically significant (t =6.249,P =0.014).The migration ability of MACF1 knockout HGC27-MACF1-/-cells was (42.48 ± 7.37)％,which was lower than that of wild type HGC27 cells ((82.35-± 4.28) ％),and the difference was statistically significant (t =5.938,P =0.017).The results of Transwell assay indicated that the number of migration cells of MACF1 knockout AGS-MACF1-/-cells was 87.0 ± 11.0,which was less than that of wild type AGS cells (200.0 ± 16.0),and the difference was statistically significant (t =5.820,P =0.028).The number of migration cells of MACF1 knockout HGC27-MACF1-/-cells was 151.0 ± 13.0,which was less than that of wild type HGC27 cells (268.5 ± 20.5),and the difference was statistically significant (t =4.840,P =0.040).The results of F-actin staining demonstrated that the number of actin filaments of MACF1 knockout AGS-MACF1-/-cells was 216.60 ± 18.09,which was less than that of wild type AGS cells (491.30 ± 5.02),and the difference was statistically significant (t =14.630,P =0.005).Conclusions The abnormally high expression of MACF1 in gastric cancer tissues may be correlated with the poor prognosis of patients with gastric cancer.MACF1 promotes the invasion and metastasis of gastric cancer cells by affecting the formation of F-actin and cell skeleton.
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Objective@#To explore the role of microtubule actin cross-linking factor 1(MACF1) in the metastasis of gastric cancer.@*Methods@#From 2009 to 2012, at The First Affiliated Hospital of Zhengzhou University, the paraffin blocks of gastric cancer and normal tissue adjacent to cancer of 107 patients who received radical gastrectomy were collected. The expression of MACF1 in tissues at protein level was detected by immunohistochemical staining. In 2017, at The First Affiliated Hospital of Zhengzhou University, fresh specimens samples of gastric cancer and normal tissue adjacent to cancer of 42 patients who received radical gastrectomy were also collected. The expression of MACF1 at mRNA level was determined by quantitative real-time polymerase chain reaction (PCR). MACF1 knockout gastric cell line was established. The effects of MACF1 on cell migration and invasion were verified by wound-healing test and Transwell assay. The effects of MACF1 on cell microtubule and actin were analyzed by filamentous actin (F-actin) staining. T-test, chi-square test and multivariate analysis were used for statistical analysis.@*Results@#The positive expression rate of MACF1 in gastric carcinoma tissues was 71.0%(76/107), which was significantly higher than that of the corresponding normal tissues adjacent to cancer (22.4%, 24/107), and the difference was significant (t=4.145, P=0.016). The expression of MACF1 at mRNA level in cancer tissues of 42 patients with gastric cancer was 6.463±0.672, which was significantly higher than that of corresponding normal tissue adjacent to cancer (1.727±0.331), and the difference was statistically significant (t=6.326, P<0.01). The differences in positive expression rate of MACF1 in different tumor infiltration depth, different TNM stage and lymph nodes metastasis were statistically significant (χ2=1.170, 7.959 and 5.288; all P<0.01). The five-year survival rate of patients with high expression of MACF1 was 32.9% (25/76), which was significantly lower than that of patients with normal MACF1 expression (64.5%, 20/31), and the difference was statistically significant (χ2=25.093, P=0.034). The high expression of MACF1 was an independent prognostic factor affecting overall survival rate in patients with gastric cancer after surgery(hazard ratio (HR)=0.513, 95%confidence interval (CI): 0.411 to 0.922, P=0.038). The results of wound-healing assay showed that at 24 hour after wound the migration ability of MACF1 knockout AGS- MACF1-/- cells was (18.77±3.82)%, which was lower than that of wild type AGS cells ((76.24±5.36)%), and the difference was statistically significant (t=6.249, P=0.014). The migration ability of MACF1 knockout HGC27-MACF1-/-cells was (42.48±7.37)%, which was lower than that of wild type HGC27 cells ((82.35±4.28)%), and the difference was statistically significant (t=5.938, P=0.017). The results of Transwell assay indicated that the number of migration cells of MACF1 knockout AGS-MACF1-/- cells was 87.0±11.0, which was less than that of wild type AGS cells (200.0±16.0), and the difference was statistically significant (t=5.820, P=0.028). The number of migration cells of MACF1 knockout HGC27-MACF1-/-cells was 151.0±13.0, which was less than that of wild type HGC27 cells (268.5±20.5), and the difference was statistically significant (t=4.840, P=0.040). The results of F-actin staining demonstrated that the number of actin filaments of MACF1 knockout AGS-MACF1-/- cells was 216.60±18.09, which was less than that of wild type AGS cells (491.30±5.02), and the difference was statistically significant (t=14.630, P=0.005).@*Conclusions@#The abnormally high expression of MACF1 in gastric cancer tissues may be correlated with the poor prognosis of patients with gastric cancer. MACF1 promotes the invasion and metastasis of gastric cancer cells by affecting the formation of F-actin and cell skeleton.
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Objective To investigate the role of cell adhesion molecule L1 like (CALL) in the genesis and development of esophageal squamous cell carcinoma (ESCC).Methods From July 2007 to December 2010,a total of 100 patients with ESCC who received radical resection of esophageal cancer were enrolled.The ESCC tissues and corresponding tumor-adjacent normal tissues were obtained.The expression of CALl was determined by tissue microarray technology and immunohistochemical staining.The CALL over-expressed esophageal cancer cell line was established.The effects of CALL on cell migration and invasion were detected by wound-healing assay and Transwell assay,respectively.The effects of CALL on actin microfilament was analyzed by filamentous actin (F-actin) staining.Chi square test,Fisher's exact test,multivariate analysis and t test were performed for statistical analysis.Results The positive expression rate of CALL in ESCC tissues was 56 ％ (56/100),which was lower than that of tumor-adjacent normal tissues (95％,95/100),and the difference was statistically significant (x2=41.114,P＜0.01).There were statistically significant differences in CALL expression at protein level among patients with ESCC of different differentiation degree,different pathological T stage,lymph node metastasis and different TNM stage (x2=13.702,5.317,21.453,Fisher's exact test;all P＜ 0.05).The five year disease related survival rate of ESCC patients with down-regulated expression of CALL was 0(0/49),which was lower than those with normal CALL expression (25.5％,13/51),and the difference was statistically significant (x2 =43.338,P＜0.01).The median survival time of CALL expression down-regulated group was 17 months,and that of normal expressed group was 38 months.CALL expression was an independent risk factor of disease special survival rate (hazard ratio (HR) 0.353,95％ confidence interval (CI) 0.188 to 0.666,P=0.001).The results of wound-healing assay showed that the migration ability of CALL overexpressed CALL-k30 cells was lower than that of Vec-k30 cells in control group on 24 hours after wound.The results of Transwell invasion test showed the number of migrating cells penetrating CALL k30 cells attached to the inferior surface of the membrane was 44.000±13.748,which was less than that of the Vec k30 cells (154.333±25.007),and the difference was statistically significant (t=5.136,P=0.036).The results of F-actin staining demonstrated that actin filaments of CALL-k30 cells was 234.667 ± 65.118,which was lower than that of Vec-k30 cells (597.000± 119.929),and the difference was statistically significant (t=4.707,P=0.042).Conclusions CALL lowers the migration and invasion abilities of esophageal cancer cells by inhibiting F-actin microfilaments.Its abnormal expression may play an important role in the genesis,development and prognosis of ESCC.
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Objective To study the pathology of primary neuroectodermal tumor (PNET),and its diagnostic standards and prognostic factors.Methods Expression of CD99,FLI-1,Syn,NSE,S-100,NF,and Vim was detected in PNET tissues stained with HE and immmunohistochemistry (En Vision method).Survival rate of 33 PNET patients with complete clinical information was analyzed with COX regression analysis method.Results The positive expression rate of CD99,FLI-1,Vim,Syn,NSE and s-100 was 88.57%,51.43%,91.42%,48.57%,45.71%,and 22.86%,respectively.The sensitivity of combined CD99 and FLI-1 was 100%.However,NF was not expressed in all PNET tissues.When other factors were unchanged,no difference was found in the effect of age on the survival rate of patients.However,a significant difference was observed in the effect of PNET site and its treatment modalities on the survival rate of patients (P