Uently expressed in the cell apex and cytoplasm of the surface mucous epithelium, and frequently but weakly in the cytoplasm of fundic and CAL120 custom synthesis pyloric glands (Figure S1 B and E), and was seen constantly at the vascular endothelium. MUC1/DF3 was NT 157 site sometimes expressed in the surface mucous epithelium, and always in the fundic glands (particularly intensely at the cell apexes), but not in the pyloric glands (Figure S1 C and F).Immunohistochemical staining of gastric adenocarcinoma. We examined gastrectomy specimens 25033180 ofResults Evaluation of Two Monoclonal Antibodies for MUCTo investigate the difference in antibody specificity between 8G7 and 1G8, we carried out RT-PCR, Western blotting and IHC analysis using two gastric cancer cell lines, SNU-16 and NCIN87 cells. The MUC4 mRNA was detected in the two gastric cancer cell lines (Fig. 1A). Consistent with the previous report [14], our data showed that 8G7 recognized a very high molecular weight protein (over 500 kD, which was the expected size for native MUC4). On the contrary, 1G8 does not show any immunoreactive bands (Fig. 1B). The same result was observed in the IHC analysis (Fig. 1C).104 early gastric cancers (pT1b2), since we wished to avoid the major degenerative changes that are frequently seen in advanced cancer tissues, and to adjust the stage for the accurate comparison between IHC findings and the clinicopathologic factors. When more than two histological types were mixed in one lesion in the gastrectomy specimens of 104 early gastric cancers, each histological pattern was evaluated independently, according to the JCGC [16]. Therefore, in the 104 gastrectomy specimens, we could evaluate 197 carcinoma foci of various histological types in total. Among the 197 lesions, there were 15 lesions of papillary adenocarcinoma (pap) (Figs. 2A ), 39 of well differentiated tubular adenocarcinoma (tub1) (Figs. 2E ), 52 of moderately differentiated tubular adenocarcinoma (tub2) (Figs. 2I ), 6 of mucinous carcinomas (muc) (Figs. 2M ), 8 of solid type poorly differentiated adenocarcinoma (por1) (Figs. 2Q ), 47 of non-solid type poorly differentiated adenocarcinoma (por2) (Figs. 2U ) and 30 of signet-ring cell carcinoma (sig) (Figs. 2Y-b), based on the context of common histological classification of gastric cancer in JCGC [16]. According to the context in the WHO classification of tumours of the stomach [17] as well as that in JCGC [16], pap and tub1 were classified into “well-differentiated adenocarcinoma”, and por1 and por2 were classified into “poorly-differentiated adenocarcinoma”. The data of the expression rate of MUC4/ 8G7, MUC4/1G8 and MUC1/DF3 were summarized in Figure 3. The detailed number and percentage of positively stained neoplastic cells using the scoring system were summarized in Table S1. Expression profile of MUC4/8G7. Among the 197 adenocarcinoma lesions, MUC4/8G7 was expressed in 83 lesions (42 ). MUC4/8G7 showed a significantly higher rate of the positive expression ( 5 of carcinoma cells stained) in well differentiated types (pap+tub1: 70 , 38/54) than that in poorly differentiated types (por1+por2: 18 , 10/55) (P,0.0001) (Fig. 3A, arrows).MUC4 and MUC1 Expression in Early Gastric CancersTable 3. Clinicopathological studies using anti-MUC4 monoclonal antibodies, 8G7 and 1G8.Organ Oral cavity Upper aerodigestive tract Salivary gland Salivary gland Salivary gland Thyroid ThyroidCarcinoma type Squamous cell carcinoma Squamous cell carcinoma Mucoepidermoid carcinoma Mucoepidermoid.Uently expressed in the cell apex and cytoplasm of the surface mucous epithelium, and frequently but weakly in the cytoplasm of fundic and pyloric glands (Figure S1 B and E), and was seen constantly at the vascular endothelium. MUC1/DF3 was sometimes expressed in the surface mucous epithelium, and always in the fundic glands (particularly intensely at the cell apexes), but not in the pyloric glands (Figure S1 C and F).Immunohistochemical staining of gastric adenocarcinoma. We examined gastrectomy specimens 25033180 ofResults Evaluation of Two Monoclonal Antibodies for MUCTo investigate the difference in antibody specificity between 8G7 and 1G8, we carried out RT-PCR, Western blotting and IHC analysis using two gastric cancer cell lines, SNU-16 and NCIN87 cells. The MUC4 mRNA was detected in the two gastric cancer cell lines (Fig. 1A). Consistent with the previous report [14], our data showed that 8G7 recognized a very high molecular weight protein (over 500 kD, which was the expected size for native MUC4). On the contrary, 1G8 does not show any immunoreactive bands (Fig. 1B). The same result was observed in the IHC analysis (Fig. 1C).104 early gastric cancers (pT1b2), since we wished to avoid the major degenerative changes that are frequently seen in advanced cancer tissues, and to adjust the stage for the accurate comparison between IHC findings and the clinicopathologic factors. When more than two histological types were mixed in one lesion in the gastrectomy specimens of 104 early gastric cancers, each histological pattern was evaluated independently, according to the JCGC [16]. Therefore, in the 104 gastrectomy specimens, we could evaluate 197 carcinoma foci of various histological types in total. Among the 197 lesions, there were 15 lesions of papillary adenocarcinoma (pap) (Figs. 2A ), 39 of well differentiated tubular adenocarcinoma (tub1) (Figs. 2E ), 52 of moderately differentiated tubular adenocarcinoma (tub2) (Figs. 2I ), 6 of mucinous carcinomas (muc) (Figs. 2M ), 8 of solid type poorly differentiated adenocarcinoma (por1) (Figs. 2Q ), 47 of non-solid type poorly differentiated adenocarcinoma (por2) (Figs. 2U ) and 30 of signet-ring cell carcinoma (sig) (Figs. 2Y-b), based on the context of common histological classification of gastric cancer in JCGC [16]. According to the context in the WHO classification of tumours of the stomach [17] as well as that in JCGC [16], pap and tub1 were classified into “well-differentiated adenocarcinoma”, and por1 and por2 were classified into “poorly-differentiated adenocarcinoma”. The data of the expression rate of MUC4/ 8G7, MUC4/1G8 and MUC1/DF3 were summarized in Figure 3. The detailed number and percentage of positively stained neoplastic cells using the scoring system were summarized in Table S1. Expression profile of MUC4/8G7. Among the 197 adenocarcinoma lesions, MUC4/8G7 was expressed in 83 lesions (42 ). MUC4/8G7 showed a significantly higher rate of the positive expression ( 5 of carcinoma cells stained) in well differentiated types (pap+tub1: 70 , 38/54) than that in poorly differentiated types (por1+por2: 18 , 10/55) (P,0.0001) (Fig. 3A, arrows).MUC4 and MUC1 Expression in Early Gastric CancersTable 3. Clinicopathological studies using anti-MUC4 monoclonal antibodies, 8G7 and 1G8.Organ Oral cavity Upper aerodigestive tract Salivary gland Salivary gland Salivary gland Thyroid ThyroidCarcinoma type Squamous cell carcinoma Squamous cell carcinoma Mucoepidermoid carcinoma Mucoepidermoid.
