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Objective: To observe the inhibitory effect of astragalus polysaccharides (APS) on growth of human breast cancer MDA-MB-231 xenograft tumor in nude mice and its effect on the apoptosis of tumor cells, in order to study the effect of APS on growth and induction of apoptosis of triple negative breast cancer MDA-MB-231 and its possible molecular mechanism. Method: Human breast cancer cell MDA-MB-231 was inoculated into the right axillary subcutaneous of BALB/c-nu female nude mice to establish the transplanted tumor model of breast cancer. Eighteen nude mice were randomly divided into 3 groups:model group (saline per day), low-dose APS group (200 mg·kg-1 APS per day), and high-dose APS group (400 mg·kg-1 APS per day), with 6 rats in each group. The drug was administered by gavage (200 μL) daily for 21 days. In the experiment, the length and diameter of breast cancer transplanted tumor were measured every two days, and the tumor volume was recorded and calculated. At the end of the experiment, the changes of tumor mass and tumor volume of the low and high-dose APS groups and the model group were observed and compared, and the tumor inhibition rate was calculated. The cell morphology in tumor tissue was observed by hematoxylin-eosin (HE) staining, and Terminal-deoxynucleoitidyl transferase mediated nick end labeling (TUNEL) was used to verify the apoptosis of breast cancer tissues. The expressions of apoptosis-related proteins, such as B-cell lymphoma/leukemia-2 protein (Bcl-2), Bcl-2 associated X protein (Bax), Caspase in tumor tissues was detected by Western blot. Result: The tumor volume of breast cancer decreased in the low and high-dose APS groups, and the tumor inhibition rates were 37.9%and 57.57%, respectively, with statistically significant differences from the model group (PP0.01). HE of tumor tissue cells showed that APS led to obvious morphological changes, with apoptosis in the tissue cells. TUNEL staining showed that the apoptosis rate of tumor cells in APS intervention groups was higher than that in control group. Western blot showed that expression of Bcl-2 protein decreased(PPPPConclusion: APS can effectively inhibit the growth of MDA-MB-231 breast cancer xenografts in nude mice and induce apoptosis in human breast cancer MDA-MB-231 cells. The mechanism may be related to the effect of APS on expressions of apoptosis-related proteins Bcl-2, Bax, Caspase-9 and Caspase-7 in breast cancer cells.
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Objective To explore the influence of Guiqiyiyuan Ointment on the expressions of caspase-3 and caspase-9 in the lung and kidney of the rats damaged by heavy ion (12C6+) radiation-induced bystander effect.Methods The Wistar male rats were equally and randomly divided into seven groups,normal control group (NCG),radiation alone group (RAG) and Chinese medicine group (CMG),with the latter two groups being redivided into 6,12 and 24h groups according to the executing time.The Chinese medicine groups were given Guiqiyiyuan Ointment by gavage for two weeks in advance.The normal control group and the radiation alone groups were given the equal normal saline.Afterwards,the right lung of the rats in the radiation alone groups and Chinese medicine groups were radiated by 2Gy 12C6+ ion once.The rats in normal control group were not radiated.All groups of rats were executed 6,12,and 24h after radiation.The protein and mRNA expressions of caspase-3 and caspase-9 in the right lung,left lung and left kidney were examined with immunohistochemistry and Q-PCR.Results Compared with the normal control group,the mRNA expressions of caspase-3 and caspase-9 in the right lung,left lung and left kidney in the radiation alone groups obviously increased 6 and 24h after radiation.While the protein expressions of caspase-3 and caspase-9 in the radiation alone group obviously increased only 24h after radiation (P＜0.01).Compared with the radiation alone groups,the expressions of protein and mRNA ofcaspase-3 and caspase-9 were obviously down-regulated in the Chinese medicine groups (P＜0.01).Conclusion By controlling the up-regulation of the expression ofcaspase-3 and caspase-9,Guiqiyiyuan Ointment can prevent the lung and kidney cell apoptosis and alleviate the damage caused by heavy ion radiation-induced bystander effect in vivo.
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This study was designed to investigate the impact of ultra-filtration extract mixture from Astragals mongholicus (UEMAM) o radiosensitivity of H22 ascitic tumor in mice to 12C6+ ions radiation. The H22 ascitic tumor model was established in mice by intraperitoneal injection of 0.2 mL H22 ascitic cells. The animals were subsequently divided into 4 groups randomly, treated with normal saline, UEMAM, heavy ion beam radiotherapy and UEMAM plus heavy ion beam radiotherapy, respectively. The body weights, abdomen circumference of the mice were measured and the mouse behavior was monitored every day; survival time was recorded to evaluate life extension effect; flow cytometry technique was used to detect H22 cell apoptosis and cell cycle; protein levels of p53, Bax, Bcl-2 and cleaved Caspase-3 were analyzed by Western blot; the single cell gel electrophoresis was used to detect the level of deoxyribonucleic acid damage (DNA damage). The results suggest that UEMAM significantly increased survival time, and decreased body weights and abdomen circumference over the saline control group. The treatment increased cell apoptosis, cycle arrest and DNA damage compared to the saline control group. UEMAM significantly enhanced the therapeutic effect of heavy ion beam radiation in survival time, and decreased body weights and abdomen circumference in the tumor-baring mice. The combination increased cell apoptosis, cycle arrest and DNA damage compared to the radiotherapy group. The results of Western blot suggest that the treatment significantly enhanced p53-induced apoptotic signals. The experiment discovered that UEMAM could improve radiosensitivity of H22 ascitic tumor through activation of p53-mediated apoptotic signal pathway.
Subject(s)
Animals , Mice , Apoptosis , Astragalus Plant , Chemistry , Cell Cycle , Cell Line, Tumor , DNA Damage , Ions , Neoplasms, Experimental , Drug Therapy , Radiotherapy , Plant Extracts , Pharmacology , Radiation Tolerance , Signal TransductionABSTRACT
To investigate the radiosensitizing effects of Radix Angelicae Sinensis-Radix Hedysari [HAS-RH [an ultra- filtration extract]] and its underlying mechanisms in human liver cancer cells H22. This study was conducted between September 2010 and August 2012 in the Gansu University of Traditional Chinese Medicine, Lanzhou, China. The groups were assigned as the control group, drug [RAS-RH] group, [12]C[6-] radiation group, and combination group. The cell counting kit-8 assay, colony formation assay, cell cycle changes, and apoptosis analysis were carried out, and survivin and casepase-9 were evaluated by reverse transcription polymerase chain reaction and Western blot analyses in the 4 groups. The inhibitory effect of RAS-RH on H22 cells was dependent on both concentration and time, RAS-RH was able to enhance the radiosensitivity of H22 cells by increasing cell survival fraction and radiosensitization parameters. Apoptosis and the gap2/mitosis [G2/M] phase transition induced by [12]C[6-] heavy ion radiation was enhanced by RAS-RH treatment. Irradiation, combined with RAS-RH, decreased survivin expression while increasing casepase-9 expression in H22 cells. The RAS-RH increased the radiosensitivity of H22 cells of [12]C[6+] heavy ion radiation significantly, and its possible mechanism of radiosensitization is to enhance caspase-dependent apoptosis through the down-regulation of survivin, thus, it can be used as an effective radiosensitizer