Expression of microtubule actin cross-linking factor 1 in gastric carcinoma and its effect on cell invasion and metastasis / 中华消化杂志

ABSTRACT

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.