Haruhiko Inufusa

Distribution of metastatic lymph nodes in colorectal cancer by the modified clearing method

PURPOSE: The aim of this study was to clarify the distribution of lymph node metastasis in colorectal cancer. We also examined the relationship between the primary tumor (T) and the regional node (N) categories of the TNM (primary tumor, regional nodes, metastasis) classification. METHOD: Lymph nodes of surgical specimens in 311 consecutive patients with colorectal cancer were studied using the modified clearing method. RESULTS: Lymph node metastasis was seen in 59.2 percent of the total cases. The upward metastasis rate was 30.7 percent. In the longitudinal spread, most of the lymph node metastasis was seen within 10 cm. On the oral side in rectal cancer, there was no metastasis beyond 4 cm. The lateral metastasis rate in rectal cancer was 8.8 percent and in the lower rectum, the rate of cancer within 6 cm from the anal verge or beyond pT3 was much higher. CONCLUSION: In the TNM classification, there was no significant difference between colon and rectal cancer except pT1 with rectal cancer. In the lower rectal cancer within 6 cm from the anal verge or beyond pT3, there is a high risk of lateral metastasis, and lateral lymph node dissection or radiation therapy should be performed.

Suppression of pulmonary metastasis using adenovirally motility related protein-1 (MRP-1/CD9) gene delivery

Previously we showed that MRP-1/CD9 might prevent tumor metastasis by suppression of cell motility and invasion of tissue barriers. The present study explored the possibility of preventing metastasis of mouse melanoma BL6 by expression of MRP-1/CD9 through gene transfer. A replication-deficient adenovirus vector was used for the in vivo transfer of MRP-1/CD9 cDNA. Intratumor injection of an adenovirus vector (rAd-MRP-1/CD9) expressing MRP-1/CD9 resulted in a 73.7% reduction in the number of pulmonary metastases of mice and the median survival time of mice treated with rAd-MRP-1/CD9 was significantly longer than those treated with the rAd-β-gal vector (103.2±8.5 days vs 71.2±5.2 days, P<0.001 respectively). These results support the expression of MRP-1/CD9 through gene transfer as a therapeutic strategy for preventing metastases and prolonging survival, and support the feasibility of gene transfer in a clinically relevant setting.

Localization of oncofetal and normal fibronectin in colorectal cancer. Correlation with histologic grade, liver metastasis, and prognosis

Background. Expression of oncofetal fibronectin (oncFN) and normal fibronectin (norFN) in colorectal cancer specimens was examined to investigate the correlation between fibronectin localization and histologic grade, liver metastasis, and prognosis.

Second-look operation for recurrent colorectal cancer based on carcinoembryonic antigen and imaging techniques

PURPOSE: The usefulness of postoperative carcinoembryonic antigen (CEA) monitoring and improvements in imaging techniques have renewed enthusiasm for second-look operations (SLO) as the most effective treatment for recurrent colorectal cancer by reresection following early detection. The aim of our study is to evaluate the role of CEA and imaging techniques-directed SLO. METHODS: Seven hundred fifty-six patients with Dukes Stages B and C, who had undergone curative resection, were monitored postoperatively using CEA and imaging techniques. An SLO was performed on any potentially resectable recurrence, and in addition, an SLO was done when a persistently rising CEA value was detected. RESULTS: Recurrence developed in 18.8 percent (142/756) of patients, and 90.8 percent (129/ 142) of the recurrences were detected within the first three years following curative resection. When comparing carcinomas of the colon with that of the rectum, the former were associated with significantly more hepatic and intra-abdominal recurrences, whereas the latter had significantly more locoregional and pulmonary recurrences. Seventy-two patients underwent SLO. Of these patients, 54.2 percent (39/72) had all of their disease resected, and 1.4 percent (1/72) had no detectable disease at the SLO. Among the 142 patients with recurrence, 71 (50 percent) patients underwent SLO. The resectable group at SLO carried a significantly better survival than the unresectable recurrence group (41.3vs.5.2 percent;P<0.01). CONCLUSIONS: Complete removal of colorectal cancer recurrences by SLO, on the basis of postoperative, follow-up CEA and imaging technique findings, results in improved survival.

Overexpression of bax Associated with Mutations in the Loop-Sheet-Helix Motif of p53

Recent investigations have revealed that mutations of the loop-sheet-helix motif of p53 is a significant factor for a poor prognosis in patients with non-small-cell lung cancer (NSCLC). To clarify this mechanism, bcl-2 and bax expression were evaluated in relation to mutations of p53. Tumor tissues of 203 patients with NSCLC were analyzed. Immunohistochemistry was performed to evaluate bcl-2 and bax expression, and polymerase chain reaction single-strand conformation polymorphism following direct sequencing was performed to investigate p53 status. A total of 79 carcinomas were bcl-2 positive, 146 carcinomas were bax positive, and 72 carcinomas had missense mutations of p53. There was no difference in bcl-2 expression in relation to p53 status. On the other hand, tumors with structural mutations of p53 had significantly lower expression of bax than those with wild-type p53 (P = 0.0026). In contrast, tumors with mutations of the loop-sheet-helix motif of p53 had significantly higher expression of bax than those with wild-type p53 (P = 0.0236). The frequency of a bcl-2/bax ratio of >/=1 was significantly lower in tumors with mutations of the loop-sheet-helix motif than that in tumors with wild-type p53 (P = 0.0240). The bcl-2/bax ratio status was a significant factor for a prognosis in patients with NSCLC (P = 0.0083). Mutations of the loop-sheet-helix motif of p53 were correlated with overexpression of bax, while other mutations of p53 were correlated with low levels of bax expression. This variation in pattern of bax expression in relation to mutant p53 might reflect the biological behavior of tumors in patients with bcl-2-positive NSCLC.

Ley glycolipid acts as a co-factor for tumor procoagulant activity

We have generated a monoclonal antibody (MAb), FS01, which inhibits the procoagulant activity (CCA‐1) produced by a human squamous cell carcinoma cell line, LK52. Expression of the antigen recognized by FS01 MAb in various cancer cell lines correlated well with the procoagulant activities of the expressing cell lines. Our objective was to characterize the molecule reacting with FS01 MAb and to analyze its involvement in the CCA‐1 procoagulant activity. The molecule was identified as a glycolipid and found to be involved in the procoagulant activity because both procoagulant activity and reactivity to FS01 MAb were lost after endoglycoceramidase treatment of CCA‐1. Furthermore, FS01 MAb recognized the Lewis Y (Ley) antigen. To confirm the involvement of a glycolipid incorporating the Ley antigen in the procoagulant activity, we attempted to purify CCA‐1 from LK52 culture supernatant. In one of the purification steps, a fraction containing low procoagulant activity (CCA‐1p) separated from the Ley‐positive fraction (CCA‐1c). Although CCA‐1c alone did not show procoagulant activity, the procoagulant activity of CCA‐1p was augmented by CCA‐1c and this augmentation was inhibited by FS01 MAb. Furthermore, CCA‐1c enhanced the procoagulant activity of 33 cell lines tested as well as CCA‐1p. In addition, purified Ley glycolipid from canine intestine augmented the procoagulant activity of CCA‐1p, and this augmentation also could be inhibited by FS01 MAb. We conclude that Ley glycolipid is a co‐factor for the procoagulant activity derived from cancer cells. Int. J. Cancer 73:903–909, 1997.

Anterior resection following posterior transsacral stapling and transection of the anal canal for low-lying rectal cancer in males

In anterior resection with anastomosis using the double-staple technique for low-lying rectal cancer in male patients, the approach to the anal canal with a stapling instrument via the abdominal area is limited by the narrow pelvis. The stapling and transection of the anal canal via the posterior transsacral approach prior to performing an anterior resection thus enables the lower rectum and anal canal to be visualized, so that the anal canal can be accurately stapled and transected even in male patients with a narrow pelvis.

Generation of a monoclonal antibody that inhibits the procoagulant activity of various cancer cell lines

Tumor procoagulant is one of the factors responsible for disseminated intravascular coagulation and metastasis. The authors found procoagulant activity in LK52 human squamous cell carcinoma cells, which they designated cancer cell‐derived blood coagulating activity 1 (CCA‐1). A monoclonal antibody (MoAb) was generated to characterize this CCA‐1 procoagulant activity. To date, antibodies that show an inhibitory effect on procoagulant activity as well as high reactivity in cancer cells are well known for their tissue factor specificity.

Enhanced urokinase-type plasminogen activator activity by extracellular matrix protein obtained from highly metastatic human lung adenocarcinoma cell line

A protein which enhanced urokinase-type plasminogen activator (u-PA) activity was purified from the extracts of extracellular matrix of highly metastatic cell line HAL-8 derived from human lung adenocarcinoma. The protein showed a single band with molecular weight of 65 kDa after the purification by Sephadex G-150 and diethylaminoethyl-cellulose followed by reversed phase separation in a high performance liquid chromatography system. The purified protein in the immobilized conditions enhanced u-PA activity in both plasminogen activation and S-2444 amidolysis by 4.6- and 2.8-fold increases in the second order rate constants (Kcat/K(m)), respectively. This protein was related to neither plasminogen nor single-chain u-PA by the immunological studies and with respect to retention time on reversed phase analysis. These results suggest that the purified material acts as an enhancer of u-PA in extracellular matrix of the cancer cells, inducing an effective tissue destruction and cell invasion and possessing a highly metastatic potential.

Characterization and Metastatic Potential of a Mouse Adenocarcinoma Cell Line XK-4 which arose in Nude Mouse during the Transplantation of Human Colon Cancer Cell Line KUMRK-4N.

新しい腺癌肝転移モデルを樹立した.1985年10月に当院で手術した直腸癌組織をヌ-ドマウスの背部に移植し, ヒト大腸癌細胞株KUM・RK-4Nを樹立し以後継代した-1990年2月にこの腫瘍より初代培養を行い培養細胞株を樹立した.この細胞をヌードマウスに尾静脈注入および前腸間膜静脈より注入したところ, 約2-3週間で肺および肝臓に明瞭な結節型の転移巣を生じた.ヒト癌でのヌ-ドマウスにおける転移はまれであり, この培養細胞の染色体分析およびアイソザイム分析ではマウス由来の癌と考えられ, また胸腺正常同系マウスへの移植は拒絶されたため, ヌードマウス由来腺癌と考えXK-4細胞と名ずけた.今回, このXK-4細胞の生物学的特性と肺および肝臓への転移能を検討した.