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To investigate the effects and mechanism of andrographolide(Andro)on type II diabetic nephropathy in Leprdb/dbmice,levels of urinary protein,serum creatinine and blood urea nitrogen after 24 h of Andro(1 mg/kg,ip)treatment on Leprdb/dbmice were measured by kit.The changes on renal pathology and sugar levels in diabetic mice treated by Andro were observed by HE staining and PAS staining.The expression of fibronectin (FN)and NOX-4 protein were analyzed by immunofluorescence.Results showed that levels of urinary protein, serum creatinine and blood urea nitrogen in Leprdb / dbmice after Andro treatment for 24 h were significantly lower than those of the control group. Kidney pathologic change was inhibited, and FN and NOX-4 expression were markedly decrease in the Andro group.In conclusion,Andro can significantly reduce the diabetic nephropathy in Leprdb/dbmice with spontaneous type II diabetes mellitus.
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Objective To investigate the role of atrial natriuretic peptide (ANP) in lung metastasis of melanoma. Methods Melanoma B16F10 cells were intravenously injected into ANP knockout mice and C57BL／6J mice. After three weeks, the mice were sacrificed, and the lungs were removed, embedded, and examined by pathology using HE staining. The numbers of metastatic foci on the lung surface and micrometastatic foci in the lung tissues were counted. Results The ANP knockout mice displayed fewer metastatic foci on the lung surface and less micrometastatic foci in the lung tissues of ANP knockout mice than in the C57BL／6J mice. Conclusions ANP deletion significantly suppresses the metastasis of melanoma in the lung of mice.
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Objective To study the effect of atrial natriuretic peptide(ANP)deletion on melanoma growth. Methods A subcutaneous xenotransplanted melanoma model was established in ANP knockout mice and C57BL/6J mice. The tumor volume was measured at the sixth day after establishment of the subcutaneous transplantation. Tumor cell proliferation and tumor-induced angiogenesis were detected by immunohistochemical staining. Results The volume and weight of xenotransplanted melanoma were significantly decreased in the ANP knockout mice compared with those in the C57BL/6J mice. The proliferative tumor cells and microvessel density were significantly decreased in the tumor tissues of ANP knockout mice compared with those in the C57BL/6J mice. Conclusions ANP deletion significantly suppresses xenotransplanted melanoma growth through inhibiting the tumor cell proliferation and angiogenesis.
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Objective To investigate the effects of P-selectin gene knockout on the biological characteristics of mice. Methods P?selectin knockout (PKO) mice and C57BL/6 mice were housed in SPF environment. To make sure that the mice were pure PKO mice by gene identification. To determine the general condition, the activity status, reproduction of P?selectin knockout mice were observed and tested. HE staining was used to observed the histological morphology and struc?tures of 6?week?old and 18?week?old P?selectin knockoutout mice. Results PKO mice and C57 mice have similar multiplica?tion, there was no significant difference in reproduction and blood routine test. The stuctures of liver, spleen, lung and kid?ney were normal. Conclusions P?selectin knockout has no significant effects on reproduction, blood routine test and major organs. This research provides animal model theoretical basis for further study on the effects of P?selection in diseases.
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[Objective] To investigate the effect of P-selectin on the intestinal tumorigenesis in the 15,24-week-old ApcMin/+ mice.[Methods] Male ApcMin/+ mice were mated with female P-selectin knockout (P-sel-/-) mice to generate AMin/+Ps-/-mice.The diameters and numbers of the intestinal tumors in the intestines of ApcMin/+ and AM/+ps-/-mice were measured using an inverted microscope.The tumor volumes were measured using the following equation:volume =0.52 × (length × width2).[Results] There were no significant difference between the volume and number of intestinal tumor in 15-week-old ApcMin/+ mice and 15-week-old A+ P-mice.The volume and number of intestinal tumor were significantly increased in 24-week-old A+P-mice compared to 24-weekold ApcMin/+ mice.P-selectin deletion significantly prolonged the survival time of ApcMin/+ mice.[Conclusions] P-selectin deletion promotes the intestinal tumorigenesis in the 24-week-old ApcMin/+ mice.
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Objective To investigate the inhibitory effect of Lycium barbarum polysaccharide ( LBP) on the tumor growth and metastasis in MMTV?PyMT mouse model of breast cancer. Methods The population of MMTV?PyMT trans?genic mice was expanded and identified. 8?week old MMTV?PyMT?positive female mice were randomly divided into LBP group and control group, 8 mice in each group. The mice of LBP group were given LBP treatment (50 mg/kg, i. p. ), and the control group was given normal saline in the same volume, once every 2 days for 4 weeks. The tumor size was measured every two days. The mice were killed at 4 weeks after treatment, the lungs were removed and fixed in Bouin′s solution to observe the number of metastatic nodules, and tumor tissues were used for immunohistochemical examination of tumor cell proliferation and vascular density. Results The tumor formation rate was 100% in the MMTV?PyMT?positive mice. The tumor weight of LBP group was 4?208 ± 0?4463 g, significantly lower than the 6?477g ± 0?3724 g in the control group (P<0?005). The number of pulmonary nodules of the LBP group was 12 ± 1?155, significantly less than that of the control group (20 ± 2?745) (P<0?05). The immunohistochemical examination using Ki67 and CD31 staining showed that tumor cell proliferation and microvessel density of the LBP group were significantly less than the NS group. Conclusions LBP inhibits breast cancer growth and metastasis through the inhibitory effect on tumor growth and metastasis, inhibition of tumor cell proliferation and angiogenesis in MMTV?PyMT mice. These mice can be used as an ideal model for studies on antitu?mor drug development for the treatment of breast cancer lung metastasis.
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<p><b>OBJECTIVE</b>To investigate the role of type I collagen as an autocrine protein in promoting the growth lung cancer cells in a three-dimensional (3D) culture system.</p><p><b>METHODS</b>Immunohistochemistry and RT-PCR were used to detect the expression of type I collagen in lung cancer specimens and 5 lung cancer cell lines. The lung cancer cell lines in 3D cultures were treated with vectors harboring short hairpin RNA (shRNA) targeting type I collagen, and the cell growth suppression was evaluated using MTT assay and colony formation assay. The lung cancer cells stimulated with supernatant from lung cancer-derived fibroblasts were tested for the expression of type I collagen mRNA.</p><p><b>RESULTS</b>Type I collagen expressions were detected in both the lung cancer tissues and the cell lines. In the 3D culture system, the growth of the cancer cell lines was obviously suppressed by shRNA-mediated type I collagen knockdown evidenced by lowered cell growth rate and colony formation ability. Stimulation with fibroblast culture supernatant resulted in enhanced expressions of type I collagen in the cancer cell lines.</p><p><b>CONCLUSION</b>Autocrine of type I collagen I is required for maintaining lung cancer cell growth in 3D cultures, and its expression is regulated by fibroblasts. These findings provide new insights into the role of type I collagen I in the tumor microenvironment and point a new direction for targeted therapy of tumors.</p>
Subject(s)
Humans , Cell Cycle , Cell Line, Tumor , Cell Proliferation , Collagen Type I , Bodily Secretions , Fibroblasts , Immunohistochemistry , Lung Neoplasms , Metabolism , RNA, Messenger , RNA, Small InterferingABSTRACT
<p><b>OBJECTIVE</b>To study the role of Slit-Robo signaling in the proliferation of human mucoepidermoid carcinoma Mc3 cells.</p><p><b>METHODS</b>We measured the expressions of Slit2 and Robo1 proteins in human mucoepidermoid carcinoma Mc3 cells using immunohistochemistry and flow cytometry. To test the role of Slit-Robo signaling in the proliferation of the cells, we treated the cells with the monoclonal antibody R5 against Robo1 receptor extracellular domain and observed the changes in the cell proliferation by cell counting and colonogenic assay; the expression of proliferating cell nuclear antigen (PCNA) in the cells following the treatment was measured with flow cytometry.</p><p><b>RESULTS</b>Treatment of Mc3 cells with the monoclonal antibody R5 caused significantly suppressed cell growth and proliferation and obviously lowered the expression of PCNA.</p><p><b>CONCLUSION</b>Slit-Robo signaling can suppress the proliferation of Mc3 cells possibly by up-regulating the expression of PCNA.</p>