Masahide Tokunou

c-MET Expression in Myofibroblasts: Role in Autocrine Activation and Prognostic Significance in Lung Adenocarcinoma

Hepatocyte growth factor (HGF) plays important roles in tumor development and progression. It is currently thought that the main action of HGF is of a paracrine nature: HGF produced by mesenchymal cells acts on epithelial cells that express its receptor c-MET. In this investigation, we explored the significance of c-MET expression in myofibroblasts, both in culture and in patients with lung adenocarcinoma. We first showed that human myofibroblasts derived from primary lung cancer expressed c-MET mRNA and protein by reverse transcription-polymerase chain reaction and Western blot analysis. Proliferation of myofibroblasts was stimulated in a dose-dependent manner by exogenously added recombinant human HGF whereas it was inhibited in a dose-dependent manner by neutralizing antibody to HGF. The addition of HGF in the culture medium stimulated tyrosine phosphorylation of c-MET. The c-MET protein was immunohistochemically detected in myofibroblasts in the invasive area of lung adenocarcinoma. Finally, the prognostic significance of c-MET expression in stromal myofibroblasts was explored in patients with small-sized lung adenocarcinomas. c-MET-positive myofibroblasts were observed in 69 of 131 cases (53%). A significant relationship between myofibroblast c-MET expression and shortened patient survival was observed in a whole cohort of patients including all pathological stages (two-sided P: = 0.0089 by log-rank test) and in patients with stage IA disease (two-sided P: = 0.0019 by log-rank test). These data suggest that the HGF/c-MET system constitutes an autocrine activation loop in cancer-stromal myofibroblasts. This autocrine system may play a role in invasion and metastasis of lung adenocarcinoma.

Altered expression of the ERM proteins in lung adenocarcinoma

Radixin is a member of the ERM (ezrin/radixin/moesin) protein family that is proposed to function as a membrane-cytoskeletal linker. Using differential display analysis, we have identified radixin as a gene down-regulated in primary lung adenocarcinoma. Real-time quantitative reverse transcription polymerase chain reaction confirmed that radixin mRNA was decreased, both in 10 early-stage bronchioloalveolar carcinomas and in 16 invasive lung adenocarcinomas, by 69% (p = 0.0002) and 82% (p < 0.0001), respectively, compared with 9 nontumor lung tissues. Similarly, moesin and ezrin mRNA levels were reduced in lung adenocarcinoma. Immunohistochemistry confirmed that cancer cells expressed very little radixin and moesin, whereas non-neoplastic alveolar and bronchiolar epithelial cells, and endothelial cells, including those within the tumor stroma, were consistently positive for these two proteins. Ezrin was localized in the apical surface of non-neoplastic bronchiolar and alveolar epithelial cells and, in contrast to radixin and moesin, the majority of tumor cells retained expression of ezrin. Localization of ezrin was altered in a significant proportion of tumor cells: whereas tumor cells forming lumina displayed membranous staining on the apical side, tumor cells with disorganized structures were either negative or diffusely positive for ezrin in the cytoplasm. Furthermore, a fraction of tumor cells invading the stroma in a scattered manner were strongly positive for ezrin. In conclusion, expression of radixin and moesin is down-regulated in lung adenocarcinoma, including early-stage bronchioloalveolar carcinoma. An intriguing implication of this finding is that these two genes may function as tumor suppressors in lung adenocarcinoma oncogenesis. Although structurally related to radixin and moesin, ezrin may have a distinct function in tumor-cell invasion.

Results of surgery for bronchogenic carcinoma located in the aortic window

Because of its critical location, lung cancer located in the aortic window can cause complications affecting the pulmonary artery trunk, aortic arch and esophagus. The results of surgical treatment are poor; however, there are few long-term survivors. In an attempt to define the indications for extended surgery, we evaluated eleven patients with non-small cell lung cancer. The tumors were classified according to their clinical extent of invasion as Type A (invading the anterior mediastinum including the central part of the pulmonary artery), Type B (invading upwardly to the mediastinum through the aortic window) or Type C (invading the posterior mediastinum including the thoracic aorta or esophagus). In the five patients with type A invasion, no metastases to the upper mediastinal lymph nodes other than the subaortic lymph nodes were found. The three patients with type B invasion had many metastases to the upper mediastinal lymph nodes. There were no metastases in the upper mediastinum in any of these patients with type C invasion, but metastases were found in a lower mediastinal lymph node #9, and a carinal lymph node. Each group clearly demonstrated a different site of mediastinal lymph nodes metastasis. The long term result was good in Type A invasion, in contrast to Type B or C invasion. Our classification may be useful for planning ones surgical approach to advanced lung cancer of the aortic window.