Razi Journal of Medical Sciences

fa مطالعه اثر درون تن و برون تن آلکالوئید کلشی سین بر رده سلول‌های سرطان پستان موشی (4T1) In Vivo and In Vitro Effect of Colchicine on Mouse Breast Cancer Cell Line (4T1) ایمونولوژی Immunology پژوهشي Research <div style="text-align: justify;">زمینه و هدف: کلشی سین یک آلکالوئید تروپولانی است که به طور معمول برای درمان نقرس استفاده می‌شود. همچنین تأثیر درمانی بر سرطان از طریق القای آپوپتوز دارد، اما مسیرهای درگیر در این فرایند مبهم باقی مانده اند. آلکالوئیدها ترکیبات مهمی هستند که پتانسیل بالایی در درمان بیماریها از جمله انواع تومور دارند. آلکالوئیدهای مختلف، اثرات ضدتکثیری و ضدمتاستازی بر روی انواع مختلف سرطان در شرایط درون تن و برون تن (in vitro و in vivo) دارند. ترکیباتی مانند کامپتوسین، وین بلاستین و کلشی‌سین در حال حاضر با موفقیت به داروهای ضد سرطان توسعه یافته اند. تحقیقات گسترده نشان داده است که کلشی سین دارای خواص ضد تکثیر و ضد سرطان در طیف گسترده ای از رده های سلولی و حیوانات است. علاوه بر این ، مطالعات اخیر نشان داده است که کلشی سین به عنوان عوامل ضد سرطان، می تواند به طور موثری باعث ایجاد آپوپتوز شود. این امر با تأثیرات بازدارنده آن بر رشد تعدادی از رده های سلولی تومور در شرایط درون تن و برون تن (invitro و in vivo)، از جمله سلول معده، کارسینوم سلول های کبدی، سلول کبدی  و کلانژیو کارسینوم نشان داده می شود. کلشی سین در دوزهای بالا بسیار سمی است که استفاده از آن را در درمان های درمانی سلول های انسان محدود می کند. فعال سازی خاص آپوپتوز در سلول‌های توموری یک رویکرد امیدوار کننده برای درمان سرطان می باشد. مطالعه حاضر به منظور بررسی برون تن و درون تن اثر این دارو بر رده سلول‌های سرطان پستانی موش انجام پذیرفت. روش کار: این  پژوهش از نوع بنیادی بوده که به روش تجربی انجام شد. ابتدا کشت سلول&lrm;های سرطان پستان موش (4T1) خریداری شده از انستیتو پاستور ایران انجام گرفت. سپس اثر کلشی&lrm;سین بر آپوپتوز و مسیرهای سیگنالینگ مرتبط، با استفاده از آزمایش های مختلف شامل سنجش زنده ماندن سلول، ریل تایم پی سی آر، رنگ آمیزی آنکسینV و پروپیدیوم یوداید، و تجزیه و تحلیل وسترن بلات تعیین شد. مطالعات درون تن نیز پس از القاء تومور در موش ها و سپس درمان آن ها انجام گرفت. یافته ها: نتایج نشان داد که کلشی&lrm;سین در غلظت 300 میکروگرم در میلی لیتر 50% سلول های سرطانی 4T1 را از بین برد. کلشی‌سین به مقدار قابل توجهی بیان mRNA ژن Bax و P53 را افزایش و مقدار بیان ژن Bcl-2   را کاهش داد. در مطالعه درون تنی نشان داده شد که در بافت توموری موش های تیمار شده با کلشیسین شواهد آپوپتوز و ناحیه تخریب و نکروز، به طور معنی‌داری وسیع تر از گروه تیمار نشده است.</div> <div style="text-align: justify;">Background & Aims: Alkaloids are important compounds that have a high potential in the treatment of diseases, including types of tumors. Different alkaloids have antiproliferative and antimetastatic effects on different types of cancer in vitro and in vivo. Compounds such as camptothecin, vinblastine, and colchicine have already been successfully developed into anticancer drugs. Colchicine binds to tubulin and interferes with tubulin polymerization, thereby disrupting microtubule formation. This issue leads to inhibiting the migration of leukocytes and other inflammatory cells and inhibiting the mitosis of dividing cells. Colchicine can also increase free cellular tubulin to limit mitochondrial metabolism in cancer cells through the inhibition of mitochondrial membrane voltage-gated anion channels. In addition, extensive research has shown that colchicine has anti-proliferative and anti-cancer properties in a wide variety of cell lines and animals. In addition, recent studies have shown that colchicine, as an anticancer agent, can effectively induce apoptosis. This is shown by its inhibitory effects on the growth of several tumor cell lines in vitro and in vivo, including gastric cell, hepatocellular carcinoma, hepatocellular carcinoma, and cholangiocarcinoma. To Colchicine is highly toxic at high doses, which limits its use in human cell therapy. Apoptosis is a regulated and programmed cell death process that involves the activation of various molecules to initiate cell death. Specific activation of apoptosis in tumor cells is a promising approach for cancer treatment. Considering the role of colchicine in inducing apoptosis, it can be considered a suitable candidate for investigating anticancer effects in the form of in vitro studies on breast cancer cell models and in vivo studies on mouse models with mammary tumors. However, the specific mechanisms of colchicine-induced cytotoxicity are controversial due to the variability of signaling pathways in different cell types. In addition, colchicine is highly toxic at high doses and the exact mechanism of its apoptotic effects in breast cancer cells is still unclear. Colchicine is an alkaloid widely used to treat gout. It also has a therapeutic effect on cancer through the induction of apoptosis, but the pathways involved in this process remain unclear. The present study was conducted to investigate the effect of this drug on mouse Breast cancer cell line in vitro and in vivo. Methods: This research is of a fundamental type that was carried out experimentally. First, mouse mammary cancer cells (4T1) purchased from Pasteur Institute of Iran were cultured. Then, the effect of colchicine on apoptosis and related signaling pathways was determined using various tests including cell viability assay, real-time PCR, annexin V and propidium iodide staining, and western blot analysis. In vivo studies were also performed after tumor induction in mice and then their treatment.Effect of colchicineand related signaling pathways were determined using different tests including cell viability assay, real-time PCR, annexin V and propidium iodide staining and western blot analysis. In vivo studies were investigated through tumor induction with 4T1 cells in mice and then their treatment. After tumor formation, the antitumor activity of colchicine was compared in cancer mice. Tumor tissues after isolation, fixation and staining with hematoxylin and eosin were examined with a microscope for the presence of cancer symptoms. Results: The results showed that colchicine at a concentration of about 300 μg/ml killed 50% of 4T1 cancer cells. Colchicine significantly increased Bax and P53 mRNA expression and decreased Bcl-2 gene expression. In the in vivo study, it was shown that in the tumor tissue of mice treated with colchicine, the evidence of apoptosis and the area of destruction and necrosis was much wider than that of the untreated group. The in vivo study showed that in the group treated with colchicine, the tumor tissue showed more evidence of apoptosis than the control group, and in the treated group, the area of destruction and necrosis was much wider than in the untreated group. In the tumor tissues of mice in the control group (without treatment), the signs of malignancy were more severe than in the group that had been treated, and abnormal mitotic forms were abundantly seen. Also, no metastases to the liver and spleen were observed in the treated group as well as in the control group. Finally, it can be concluded that the oral intake of pure colchicine reduces the size of induced tumors in mice, prevents further proliferation and metastasis, and induces Apoptotic. Studies have shown that tumors that grow faster, such as tumors from 4T1 cancer cells, are more invasive and metastasize to the surrounding tissues, but the cells of these tumors (with a high proportion of dividing cells) are the most sensitive to toxic compounds of the cell cycle, such as colchicine. They have sin. Continuous use of these compounds causes a high percentage of exposed cells to be destroyed. Reduction in the size of tumors and metastasis of cells to other places in mice treated with colchicine was observed in the present study. Conclusion: Studies have shown that tumors that grow faster, such as tumors from 4T1 cancer cells, are more invasive and metastasize to the surrounding tissues, but the cells of these tumors (with a high proportion of dividing cells) are the most sensitive to They have cell cycle toxic compounds such as colchicine. Continuous use of these compounds causes a high percentage of exposed cells to be destroyed. Reduction in the size of tumors and metastasis of cells to other places in mice treated with colchicine was clearly observed in the present study. Colchicine had an effective role in inhibiting the proliferation of breast cancer cells, and its oral use reduced the size of induced tumors in mice, prevented further proliferation and metastasis, and induced apoptosis. The mechanism of toxicity of colchicine for cancer treatment is still not clear enough. In the present research, the results of all the tests showed that colchicine has an effective role in inhibiting the proliferation of mouse breast cancer cells and apoptosis through increasing the expression of pro-apoptotic genes and proteins Bax and p53, decreasing the expression of anti-apoptotic Bcl2 genes and proteins, and ultimately induces the activation of caspases.</div> کلشی سین, سلول سرطان پستان موش, آپوپتوز, مطالعات درون و برون تن Colchicine, Mouse Breast Cancer cell line, Apoptosis, in vivo and in vitro study 1 16 http://rjms.iums.ac.ir/browse.php?a_code=A-10-7506-1&slc_lang=fa&sid=1 Elham Adham Foumani الهام ادهم فومنی 3900319475328460080036 3900319475328460080036 No PhD Student of cell and Molecular biology, Department of Biology, Science and Research Branch, Islamic Azad University, Tehran, Iran دانشجوی دکتری تخصصی زیست شناسی سلولی و مولکولی، گروه زیست‌شناسی، واحد علوم و تحقیقات تهران، دانشگاه آزاد اسلامی، تهران، ایران Shiva Irani شیوا ایرانی 3900319475328460080037 3900319475328460080037 No Associate Professor, Department of Biology, Science and Research Branch, Islamic Azad University, Tehran, Iran دانشیار، گروه زیست‌شناسی، واحد علوم و تحقیقات تهران، دانشگاه آزاد اسلامی، تهران، ایران Yalda Shokoohinia یلدا شکوهی نیا 3900319475328460080038 3900319475328460080038 No Associate Professor, Department of Pharmacognosy and Biotechnology, School of Pharmacy, Kermanshah University of Medical Sciences, Kermanshah, Iran دانشیار، گروه فارماکوگنوزی، دانشکده داروسازی، دانشگاه علوم پزشکی کرمانشاه، کرمانشاه، ایران Ali Mostafaie علی مصطفایی amostafaie@kums.ac.ir 3900319475328460080039 3900319475328460080039 Yes Professor, Medical Biology Research Center, Kermanshah University of Medical Sciences, Kermanshah, Iran استاد ایمونولوژی، مرکز تحقیقات بیولوژی پزشکی، دانشگاه علوم پزشکی کرمانشاه، کرمانشاه، ایران