Increased survival of rat hepatocytes in serum-free medium by inhibition of a trypsin-like protease associated with their plasma membranes

Nakamura, Toshikazu; Asami, Osamu; Tanaka, Keiji; Ichihara, Akira

doi:10.1016/0014-4827(84)90769-9

Cited by 48 publications

(14 citation statements)

References 24 publications

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“…McKeehan et al (1986) indeed showed that a,-AT exhibits growth-stimulatory activities on endothelial cells. Nakamura et al (1984) have also shown that trypsin inhibitors including a,-AT increase the survival of rat hepatocytes in serum-free culture by inhibiting trypsin-like proteinase associated with plasma membrane. The tumor a,-AT may therefore act as an endothelial cell growth factor to maintain blood circulation in the tumor and/or as an autocrine cell survival factor by regulating the excess proteinase activities of tumor cells themselves.…”

Section: Discussionmentioning

confidence: 94%

Prognostic significance of alpha‐1‐antitrypsin in early stage of colorectal carcinomas

Karashima

Kataoka

Itoh

et al. 1990

Intl Journal of Cancer

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We have previously shown that human colorectal carcinoma cell lines, RCM-1 and CoCM-1, synthesize alpha-1-antitrypsin (alpha 1-AT) in culture. We have studied immunohistochemically the incidence of alpha 1-AT on histologic sections from paraffin-embedded tissues of surgically resected colorectal carcinomas and their metastatic foci, polypectomized adenomas, and normal mucosae. alpha 1-AT was detected in 89 (61%) of 145 carcinomas (including 14 carcinomas in adenoma), and 12 (39%) of 31 adenomas. But only 2 (4%) of 55 normal colorectal mucosae were positive for alpha 1-AT. In metastatic tumor cells of colorectal carcinomas in lymph nodes and other organs, alpha 1-AT positivity was 60% and 82%, respectively. The incidence of alpha 1-AT was markedly higher in advanced adenocarcinomas than in early ones and more frequent in adenocarcinomas of right side (including transverse colon) than those of left side and rectum, regardless of their histological malignancy grades. In mucinous carcinomas the frequency was greater (8 of 9 cases) than in conventional adenocarcinomas. Clinical follow-up of the patients with colorectal carcinomas suggested that alpha 1-AT positivity in Dukes' stage A/B tends to correlate with unfavorable prognosis irrespective of the grade of histologic differentiation of carcinoma, but there is no significant relation in Dukes' stage C/D. Our findings suggest that alpha 1-AT in colorectal carcinoma is related to the invasive and metastatic capacity. It may thus serve as a biologic marker for prognosis of colorectal carcinomas at relatively early stages (Dukes' stage A/B).

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Section: Discussionmentioning

confidence: 94%

Prognostic significance of alpha‐1‐antitrypsin in early stage of colorectal carcinomas

Karashima

Kataoka

Itoh

et al. 1990

Intl Journal of Cancer

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“…The isolated cells were seeded at a density of 5 ϫ 10 4 cells/cm 2 and cultured in Williams' medium E (ICN Biomedicals, Costa Mesa, CA) with 10% fetal calf serum (GIBCO, Grand Island, NY), 0.9 M dexamethasone (Takeda Pharmaceutical Industries, Osaka, Japan), and 10 M Actrapid MC insulin (Novo Industri, Copenhagen, Denmark). At 2 h after seeding, the medium was replaced with serum-and hormone-free Williams' medium E containing 0.2 g/ml aprotinin (Sigma Chemical) (24).…”

Section: Animals Male Sprague-dawley Rats (Shizuoka Laboratory Animalmentioning

confidence: 99%

Rho-kinase inhibitor prevents hepatocyte damage in acute liver injury induced by carbon tetrachloride in rats

Ikeda

Kume²,

Tejima³

et al. 2007

American Journal of Physiology-Gastrointestinal and Liver Physi

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A protective effect of Rho-kinase inhibitor on various organ injuries is gaining attention. Regarding liver injury, Rho-kinase inhibitor is reported to prevent carbon tetrachloride (CCl4)- or dimethylnitrosamine-induced liver fibrosis and hepatic ischemia-reperfusion injury in rats. Because Rho-kinase inhibitor not only improved liver fibrosis but also reduced serum alanine aminotransferase (ALT) level in CCl4-induced liver fibrosis, we wondered whether Rho-kinase inhibitor might exert a direct hepatocyte-protective effect. We examined this possibility in acute CCl4 intoxication in rats. Rho-kinase inhibitor, HA-1077, reduced serum alanine ALT level in rats with acute liver injury induced by CCl4 with the improvement of histological damage and the reduction of the number of apoptotic cells. In cultured rat hepatocytes in serum-free condition, HA-1077 reduced apoptosis evaluated by quantitative determination of cytoplasmic histone-associated DNA oligonucleosome fragments with the reduction of caspase-3 activity and the enhancement of Bcl-2 expression. HA-1077 stimulated phosphorylation of Akt, and wortmannin, an inhibitor of phosphatidylinositol 3-kinase (PI3-kinase)/Akt pathway, abrogated the reduction of hepatocyte apoptosis by HA-1077 in vitro. Furthermore, wortmannin abrogated the reduction of serum ALT level by HA-1077 in rats with acute liver injury induced by CCl4, suggesting that the activation of PI3-kinase/Akt pathway may be involved in the hepatocyte-protective effect by Rho-kinase inhibitor in vivo. In conclusion, Rho-kinase inhibitor prevented hepatocyte damage in acute liver injury induced by CCl4 in rats and merits consideration as a hepatocyte-protective agent in liver injury, considering its direct antiapoptotic effect on hepatocytes in vitro.

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“…23 Although this possibility was not tested in our study, the fact that predominant molecular forms of APP expressed in carcinoma cells, as well as in SW837 cells, are the Kunitz-domain-containing isoforms 4,5 may support the possible role of the Kunitz domain in the growth of cancer cells. In fact, serine proteinase inhibitors and their proteolytic fragments have been shown to enhance the cellular growth and/or survival of endothelial cells, 24 hepatocytes 25 and fibroblasts 26 in vitro and of tumor cells in vivo. 27 An alternative possibility is that APP may modulate the cellular growth and survival indirectly via its potential roles in cell-ECM and cell-cell interactions.…”

Section: Reduced Tumor Growth Of App Antisense Clones In Nude Micementioning

confidence: 99%

Amyloid β protein precursor is involved in the growth of human colon carcinoma cell in vitro and in vivo

Jingyan¹,

Kataoka²,

Itoh³

et al. 2001

Int. J. Cancer

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Amyloid ␤ protein precursor (APP) is a membrane-bound protein ubiquitously expressed in a variety of types of cells. However, its biological functions remain largely uncertain, particularly in non-neural cells and tumors. Our previous studies revealed that a secreted form of APP having a Kunitztype inhibitor domain is a major serine proteinase inhibitor secreted by human colon carcinoma cells. In our study, we used an antisense RNA strategy to selectively inhibit the expression of APP in the human colon carcinoma cell line SW837. A vector capable of expressing an antisense mRNA complementary to 911 bases of the 5 end of APP mRNA was transfected into SW837 cells. After selection, 2 stably transfected antisense clones were obtained in which both the APP protein and mRNA were significantly suppressed. The proliferative potential and colony-forming efficiency of the antisense clones in vitro were markedly suppressed compared with the parent and mock-transfected clones. The addition of the conditioned medium of parent cells or purified secretory APP enabled these antisense effects to be overcome in vitro. The suppressed growth was also observed in vivo when the cells were injected subcutaneously into nude mice. Histologically, formation of tubular structures appeared to be suppressed in the antisense clones in vivo. These observations suggest potentially important roles of APP in cellular proliferation and differentiation of colon carcinoma cells. Key words: amyloid ␤ protein precursor; colon carcinoma cell; antisense; growthAmyloid ␤ protein precursor (APP) is a large precursor protein of ␤-amyloid found in neurite plaques of Alzheimer's disease, and it exists in multiple isoforms as a result of alternative splicing of the 19 exons encoded by the APP gene. 1 With regard to exons, exon 7 encodes for 57 amino acids with considerable homology to a Kunitz-type serine proteinase inhibitor, and the secreted protein containing the inhibitor domain is identical to proteinase nexin-II, which was initially isolated from human fibroblasts by its property of forming complexes with epidermal growth factor-binding protein. 2 APP is a ubiquitous protein and synthesized by most cells throughout the body. 1 However, its biological functions are largely unknown, particularly in non-neural tissues. Previously, we reported that human tumor cell lines consistently secreted highmolecular weight trypsin inhibitors, which were identical to the secreted form of APP (sAPP) with Kunitz domain. 3,4 Indeed, sAPP represented the major trypsin inhibitor produced by a panel of colon carcinoma cell lines. 5 This evidence suggests that APP and/or sAPP may have important, though undefined, biological functions in tumor cells, including colon carcinoma cells.To date, a limited amount of evidence is available for the possible role of APP in non-neural cells and essentially nothing is known about its function in tumors of systemic organs. 1 Saitoh et al. 6 showed that APP can be a potent growth factor for fibroblasts as well as for thyroid follicular e...

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Increased survival of rat hepatocytes in serum-free medium by inhibition of a trypsin-like protease associated with their plasma membranes

Cited by 48 publications

References 24 publications

Prognostic significance of alpha‐1‐antitrypsin in early stage of colorectal carcinomas

Prognostic significance of alpha‐1‐antitrypsin in early stage of colorectal carcinomas

Rho-kinase inhibitor prevents hepatocyte damage in acute liver injury induced by carbon tetrachloride in rats

Amyloid β protein precursor is involved in the growth of human colon carcinoma cell in vitro and in vivo

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