[Effects of human alpha-mannosidase Man2c1 transgene on growth and metastasis of transplanted tumor in mice].
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To study the effect of human alpha-mannosidase Man2c1 transgene on tumor growth and metastasis in mice.Hepatoma cell H22 or squamous epithelial carcinoma cell S180 was subcutaneously inoculated into the right armpit of mice (wild type mice and 28#, 35#, and 54# transgenic mice). Tumor size was measured every week. Mice were sacrificed on day 9 or 10 and then the tumors were exercised and weighted. Tumors and lungs were fixed in formaldehyde and sectioned. The sections were stained with hematoxylin/eosin and examined under microscope. The red blood cells in spleen were destroyed by Tris-NH4Cl. Natural killer (NK) cell activity was detected with Yac-1 cell as target.H22 and S180 tumors grew faster in all the three transgenic mice (28#, 35#, and 54#) than in wild type mice. The average size and weight of tumors between the transgenic mice and wild type mice were significantly different (P<0.05). Most tumors in the transgenic mice invaded the surrounding tissues. In contrast, nearly all the tumors in wild type mice were capsulized. Three of 10 28# transgenic mice, 5 of 10 35# transgenic mice, 3 of 10 54# transgenic mice, and 1 of 10 wild type mice showed lung metastasis of H22 tumor. Two of 6 28# transgenic mice, 3 of 6 35# transgenic mice, 1 of 6 54# transgenic mice, and 0 of 6 wild type mice showed lung metastasis of S180 tumor. No difference of NK activity in spleen cells was observed between the transgenic mice and wild type mice.hMan2c1 transgene promotes growth, invasion, and metastasis of transplanted H22 and S180 tumors in mice. hMan2cl transgene does not affect NK activity in splenocytes.Keywords:
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Summary Passage of a mouse tumor graft, Sarcoma I from A/He mice, through C57BL/6Jax mice, actively immunized with whole nonviable Sarcoma I tumor antigen, resulted in a biologic modification of the tumor graft. The modification of the tumor was evidenced by progressive growth of the altered tumor in serial passages in normal adult mice of three genetically unrelated inbred strains. Sarcoma I tissue from immunized C57BL/6Jax mice showed progressive evidence of anaplasia on being transplanted from the immunized C57BL/6Jax mice to normal C3H, C57BL/6Jax, and BALB/c mice. The tumor tended to revert to its original morphology when transplanted back to A/He mice from the subline passages in the three unrelated mouse strains. The sarcoma graft established in C3H mice, when transplanted in serial passage to normal A/He mice and then back to normal C3H mice, grew progressively, and all mice of both strains died with large tumors. A variant of Sarcoma I, designated as Sarcoma Mo, has been described.
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The role of natural killer (NK) cells in tumor growth and metastasis was studied in syngeneic normal and beige inbred C57BL/6 mice. Mice with the beige point mutation have been shown to be deficient in nonstimulated NK activity. Tumor-passaged B16 malignant melanoma cells were refractory to NK activity as determined by in vitro assay, but after in vitro culture they became sensitive to NK activity. The NK-insensitive B16 tumor grew and metastasized similarly in normal and beige mice. However, the NK-sensitive B16 tumors grew more slowly and produced fewer metastases in normal mice than in NK-deficient beige mice. Activation of NK cells by lymphocytic choriomeningitis virus infection decreased the rate of growth and number of metastases of both NK-sensitive and NK-insensitive tumors in both normal and beige mice. These results suggest the importance of NK cells as a determinant of tumor growth and metastasis.
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Adoptive Cell Transfer
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Some tumors of no mammary tissue origin (sarcoma, hepatoma) grew more effectively in MTV-free mice (C3Hf) as compared with MTV-infected mice (C3H/He), whereas the tumors of mammary gland origin grew more rapidly in the latter ones. It has been shown that C3H/He mice develop a stronger specific immune reaction against hepatoma 22a cells than do C3H/f mice and that the animals of MTV-bearing subline exhibit a higher natural killer activity against these cells.
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Inbred A/J, CBA/J, and C57BL/6 mice at age 7-8 weeks were inoculated with 1 mg urethan/g body weight. Three months later, 13.2, 1.8, and 0.1 tumors per lung were found in these mice, respectively. Cytotoxicity by spleen cells of normal and urethan-treated mice was analyzed in a 4-hour 51Cr release assay against YAC-1 cells. During the first 2 weeks after treatment, urethan strongly suppressed the cytotoxicity by spleen cells of A/J mice but had relatively little effect on the reactivity of spleen cells of CBA/J mice. Natural killer (NK) activity was not suppressed in urethan-treated C57BL/6 mice. The effect of urethan on natural in vivo antitumor resistance in the lungs was studied by measurement of elimination from the lungs of [125I]-deoxyuridine-labeled YAC-1 cells. Urethan caused profound and sustained suppression of in vivo NK reactivity in the lungs of A/J mice, had only a transient effect on CBA/J mice, and had no detectable effect on C57BL/6 mice. The differences in the effects of urethan on the natural antitumor resistance in the three strains of mice could not be attributed to a generally greater susceptibility of A/J mice to inhibition of NK activity, since cyclophosphamide (0.15 mg/g) equally suppressed the clearance from the lungs of the radiolabeled tumor cells in A/J, CBA/J, and C57BL/6 mice. These results indicate a positive correlation between susceptibility of A/J, CBA/J, and C57BL/6 mice to urethan-induced lung carcinogenesis and inhibition of in vitro and in vivo natural cell-mediated cytotoxicities and support the hypothesis that NK cells play a role in resistance to urethan-induced lung carcinogenesis.
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Both systemic and in situ natural killer (NK) activity was assessed in young and old C57BL/6N and CBA/N mice bearing primary murine sarcoma virus-induced tumors. Splenic NK activity was depressed in tumor-bearing mice relative to normal age-matched controls. In situ NK activity was detected in young mice, and the level paralleled that found in the spleens of the same mice. In older mice, in situ NK activity was not detected. Furthermore, the depressed levels of activity in the spleens of tumor-bearing mice seemed to reflect systemic effects rather than migration of NK cells from the spleen to the tumor site.
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AIM To explore the effect of immune cells in various mice with different immune on growth of human hepatocarcinoma cells in these mice and xenospecific cytotoxicity of these mice to hepatocarcinoma cells. METHODS The human hepatocarcinoma cells cultured were inoculated subcutaneous into the posterior limbs in various mice with different immune (including normal immune mice-BALB/c, B lymphocyte deficient mice-CBA/N, T lymphocyte deficient mice-BALB/c nude and T, B lymphocytes combined deficient mice-C.B-17 SCID) and their growth in the mice. Were surveyed And the spleen cells were harvested from both experiment groups in 5 weeks after being inoculated and from control groups a to examine direct cytotoxicity to the hepatocarcinoma cells. RESULTS There were no carcinoma growth in BALB/c, CBA/N mice and immunoreconstructed SCID mice with BALB/c mice peripheral blood lymphocytes (BALB/c-PBL-SCID) but 100% carcinoma growth in SICD, nude and immunoreconstructed SCID mice with CBA/N mice peripheral blood lymphocytes (CBA/N-PBL- SCID). And the spleen cells in experiment groups of BALB/c and CBA/N mice displayed strong cytotoxicity to target cells and the lymphocytes of immunoreconstructed SCID mice generated a little lysis of target cells, but those of control groups and nude, SCID mice generated no lysis of target cells. CONCLUSION Tlymphocytes which are xenospecific cytotoxic mayplay a main role of immune killing in rejection to xenografted tumor but the comprehensive immune may be necessary in the tumor immunity.
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Syngeneic mammary gland tumor (MMT-1) grew slower in non-factor C3H/f (MTV-L) mice than in wirus-positive C3H/He (MTV-S) mice. The immunization of mice with tumor (MMT-1), inoculated and later excised, revealed a higher immune reactivity in the mice of the non-factor substrain. In studying the dynamics of the cytotoxic activity of lymphocytes in regional lymph nodes cytotoxically active lymphocytes were found to appear in C3H/f mice on day 6 after the inoculation of tumor (MMT-1), while the areactive state of lymphocytes was observed in C3H/He mice on days 3 and 24 and in C3H/f mice on day 24 after the inoculation of tumor (MMT-1). Spleen lymphocytes in C3H/f mice had no cytotoxic effect on tumor cells.
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