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Objective At present, the clinical significance and biological function of Msi1 (Musashi1) in colon cancer are still not very clear. So, a comprehensive understanding of the expression and role of Msi1 in colon cancer has important clinical and theoretical significance. This study is to investigate the clinical significance of Msi1 gene and its biological role in colon cancer by lentiviral vector to interfere with Msi1 gene expression in colon cancer SW480 cells. Methods 20 colon cancer specimens were collected from the Second Surgery Department of the Fourth Hospital of Hebei Medical University from October 2013 to May 2014. Each specimen was collected from the cancer tissue and the adjacent intestinal wall tissue. Western blot was performed to determine the protein expression of Msi1 in tumor tissues and adjacent normal tissues from colon cancer patients. The relationship between Msi1 protein expression and clinical characteristics was further analyzed. The lentiviral vector was used to construct a stable SW480 cell line with low expression of Msi1. The lentivirus containing two different interference sequences (shmsi1-1 and shmsi1-2) was transfected into the target cells, and the colon cancer cells were divided into control group (without any treatment), shMsi1-1 group (transfected shMsi1-1) and shMsi1-2 group (transfected shMsi1-2). The two lentivirus silencing effects were detected by Western blot. Cell proliferation was detected by CCK-8 assay, Clone formation assay was conducted to detect the colony forming ability, and Flow cytometry analysis was used to examine the apoptosis rate. Results The protein expression of Msi1 in colon cancer tissue(0.863±0.208) was significantly higher than that in adjacent normal tissues(0.272±0.078), and the difference was statistically significant(P<0.001). The relative expression of Msi1 protein in shMsi1-1 and shMsi1-2 groups (0.299±0.111 and 0.207±0.087) was significantly lower than that in the control group (1.000±0.149) (P<0.001). The proliferation rate of shMsi1-1 and shMsi1-2 at 48 h and 72 h was significantly lower than that of the control group (P<0.01). Compared with the control group (296.33±64.04), shMsi1-1 group (92.00±43.31) and shMsi1-2 group (78.67±32.87) were significantly decreased (P<0.01). Compared with the control group [(4.01±0.26) %], the apoptosis rate of shMsi1-1 group, shMsi1-2 group [(10.22±1.04) %, (10.87±1.27) %] was significantly increased (P<0.001). Conclusion Interference with Msi1 gene expression inhibits proliferation of colon cancer SW480 cells and promotes tumor cell apoptosis. This finding provides a new intervention target for the clinical treatment of colon cancer.
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Objective To investigate the relationship between the expression of linc-VLDLR in extracellular vesicles (EVs) and the development and drug resistance of esophageal carcinoma.Methods Fifty percent of inhibitory concentration (ICs0) ofadriamycin (ADM) for Eca109 cells was detected by MTT assay,after the treatment of esophageal squamous cell carcinoma Eca109 cell line with different concentrations of ADM for 24h.The culture medium was treated with 3 concentrations of ADM based on the IC50 for 24h for extracting EVs in Eca109 cells.linc-VLDLR mRNA expression in EVs was detected by qRT-PCR assay.ICs0 of ADM for Eca109 cells intervened by EVs for 48h was detected by MTT assay.Cell cycle was detected by FCM and linc-VLDLR and ABCG2mRNA expressions in Eca109 cells were detected by qRT-PCR after the treatment of the EVs for 48h.Results ICs0 of ADM acting on Eca109 cells for 24h was 0.44 ± 0.02μg/ml,so ADM concentrations of 0.2,0.4,0.8μg/ml were choosed in the following studies.EVs were extracted from the supernatant after the treatment of 0,0.2,0.4,0.8μg/ml ADM for 24h and were labeled as EVs1,2,3 and 4 respectively.LincVLDLR mRNA expression in EVs4 was significantly higher than that in EVs1-3 (P＜0.01).ADM ICs0 for Eca109 cells in EVs4 group was significantly higher than that in other groups after the treatment of EVs1-4 on Eca109 cells for 48h (P＜0.05).Flow cytometry results showed that the proliferation index of Eca109 cells in EVs4 group was significantly higher than that in EVs 1-3 and control groups (P＜0.01).Linc-VLDLR and ABCG2 mRNA expression levels in Eca109 cells of EVs4 group were significantly higher than these of EVs1-3 and control groups (P＜0.05).Conclusions High expression of linc-VLDLR and ABCG2 gene in esophageal cancer cells is involved in the formation of esophageal cancer resistance.EVs released by drug-resistant cells can upregulate the expression of ABCG2 in esophageal cancer cells and regulate the drug resistance of esophageal cancer cells,which is related to the linc-VLDLR gene carried by EVs.
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Objective To study the inhibitory effect of the Chinese medicine on S180 tumor-bearing mice and the effect on the immune function of the mice by respiratory drug administration. Methods The S180 cells were inoculated into the abdominal cavity of Kunming mice. After the ascites was formed, the left anterior limb of mice was inoculated with ascites to establish the subcutaneous transplantation tumor model. Drug was administrated when the tumor volume reached 0.06cm3. The experimental grouping and intervention measures are as follows (n=6): mice in control group received no intervention, and fed normally. Mice in experiment group were treated with respiratory tract inhalation of Chinese medicine gas, daily administration 6h, once a day, continuous administration of 21 days. Mice in adriamycin (ADM) group were intraperitoneally injected 1mg/kg of ADM, once a week, a total of 3 times. General signs of the mice including activity and eating status were observed daily. The body weight of mice was measured every other day. Blood samples were taken 2 days after stopping drug inhalation by inner canthus vein bleeding for detection of CD3+, CD4+ and CD8+ T lymphocyte ratio by flow cytometry. The subcutaneous nodule was dissected and weighed, and some tumor tissues were pathologically observed. Part of tumor tissues was prepared into single cell suspension to detect the cell apoptosis by flow cytometry. Results The CD3+, CD4+ T lymphocytes ratio and the ratio CD4+/CD8+ were significantly higher in experiment group than in control group (P0.05). The apoptotic rate increased significantly in experiment group than in control group (P<0.05). Histopathological observation showed that, in control group, the transplanted tumor tissue had rich cells arranged closely with necrosis limited; while in the experiment group the necrosis of tumor tissue distributed widely without obvious regional, the fibrous tissue proliferated, the tumor cells scattered in the arrangement with peritumoral lymphocytic infiltration. More necrotic tissues were also observed in ADM group. Conclusion Respiratory tract inhalation of Chinese medicine may inhibit the growth of S180 subcutaneous transplanted tumor in mice to a certain extent.
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Many epidemiologic and clinical studies have indicated that the frequency of breast cancer was lower in parous women than in nulliparous women. Moreover, the incidence of breast cancer has been reported to be lower in women with early childbirth than in women with late childbirth. To verify the effect of childbirth and the age at first childbirth on carcinogenesis and progression of breast cancer, we induced breast cancer by 7,12-dimethylbenanthracene (DMBA) in 120 female Sprague-Dawley (SD) rats, and divided them into control or experimental (DMBA-treated) nulliparous, early childbirth, and late childbirth groups to observe the incidence, latency, and size of breast cancer. Argyrophilic nucleolar organizer regions (AgNOR) count and the expression of C-erbB-2, proliferating cell nuclear antigen (PCNA), Ki-67, and minichromosome maintenance protein 2 (MCM2) in breast cancer tissues were detected by immunohistochemistry. The breast cancer incidences were 95.0%, 16.7%, and 58.8% in the experimental nulliparous, early childbirth, and late childbirth groups, respectively (all P < 0.05). Between any two of these groups, the latency was significantly different, but tumor size was similar. AgNOR count and the expression of C-erbB-2, PCNA, Ki-67, and MCM2 were significantly higher in the experimental nulliparous group than in the experimental early or late childbirth groups (P < 0.05), but no significant differences were observed between the latter two groups. Taken together, the results suggest that childbirth, especially early childbirth, can reduce the incidence and postpone the onset of DMBA-induced breast cancer.