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Objective:To investigate the effect of lentivirus-mediated silencing of mitochondrial ribosomal protein L35 (MRPL35) gene on the growth of human esophageal cancer TE-1cells, and to clarify its mechanism.Methods:Three kinds of human esophageal cancer cells, TE-1, ECA109and KYSE150, were selected.The relative expression levels of MRPL35mRNA in three kinds of cells by real-time quantitative PCR.The esophageal cancer TE-1cells were divided into shMRPL35group and shCtrl group, and the cells were infected with si-RNA lentivirus and si-RNA lentivirus;the esophageal cancer cell line stably silenting the MRPL35gene was established.Real-time quantitative PCR and Western blotting methods were used to detect the efficiency of MRPL35gene silencing.The cell growth curves in various groups were detected by CCK-8method, and the apoptotic rates were detected by flow cytometry after AnnexinⅤ-PE/7AAD double staining.Results:Three kinds of esophageal cancer cells expressed MRPL35gene, and the expression levels were not statistically significant between them (P＞0.05) .The results of real-time quantitative PCR and Western blotting methods showed that the mRNA and protein levels of MRPL35in the TE-1cells in shMRPL35group were significantly lower than those in shCtrl group (P＜0.05) .Compared with shCtrl group, the cell growth speed in shMRPL35group was decreased (P＜0.05) , and the apoptotic rate was significantly increased (P＜0.01) .Conclusion:Silencing MRPL35gene can inhibit the proliferation of esophageal cancer TE-1cells and plays a role through the apoptotic pathway.
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Objective To study the efficacy of L-3,5-diiodotyrosine (DIT) and inorganic iodine (KIO3) in iodine-deficiency Wistar rats.Methods Sixty Wistar rats,weighting about 160-180 g,were divided into two groups according to body weight by the random number table method:iodine-deficiency model (40 rats) was fed with low-iodine food (the iodine content was 35.9 μg/kg);optimal-iodine model (20 rats) was fed with low-iodine food and given with KIO3 water (the iodine content was 18 mg/L) 0.5 ml by intragastric once a day.Model was established for 3 months.Iodine-deficiency model was subdivided into low iodine (LI) group,KIO3 group and DIT group,eight,nine,ten rats in each group;from optimal-iodine model,nine rats were randomly selected as optimal iodine (NI) group.LI group was fed with low-iodine food;KIO3 group was fed with low-iodine food and given with KIO3 water (the iodine content was 18 mg/L) 0.5 ml by intragastric once a day;DIT group was fed with low-iodine food and given with DIT water (the iodine content was 18 mg/L) 0.5 ml by intragastric once a day;NI group was fed with low-iodine food and given with KIO3 water (the iodine content was 18 mg/L) 0.5 ml by intragastric once a day.After 3 months,24-hour urine of the rats was collected.According to the method for determination of iodine in urine by As3 +-Ce4+ catalytic spectrophotometry (WS/T 107-2006),iodine content in urine was detected.Rats were anesthetized intraperitoneally with 25％ urethane,blood from abdominal aortic was collected to determinate the serum thyroid hormone [total triiodothyronine (TT3),total thyroxine (TT4),free triiodothyronine (FT3),free thyroxine (FT4)] level in rats by automatic electrochemical luminescence immunoassay.All the rats were sacrificed to analyze the thyroid weight.Results ① The urine iodine showed significant differences in the four groups (x2 =25.24,P ＜ 0.05).The median of urine iodine concentration in the LI,NI,KIO3 and DIT groups were 3.00,286.14,223.37,214.33 μg/L,respectively.The urine iodine concentration in LI group was significantly lower than those of other three groups (all P ＜ 0.05).② The serum TT3,TT4,FT3,FT4 levels showed significant differences in the four groups (F =63.48,140.73,130.20,365.27,all P ＜ 0.05).And the hormone levels in KIO3 group were lower than those of the DIT group [TT3:(1.57 ± 0.20) vs.(1.97 ± 0.18) mmol/L,TT4:(51.23 ± 4.90) vs.(71.94 ± 5.27) mmol/L,FT3:(5.34 ± 0.45) vs.(6.98 ± 0.33) pmol/L,FT4:(26.18 ± 2.30) vs.(35.47 ± 2.28) pmol/L,all P ＜ 0.05].③The color of thyroid in KIO3 and DIT groups became pale pink.The absolute and relative thyroid weight showed significant differences in the four groups (F =225.05,345.40,all P ＜ 0.05).The absolute thyroid weight [(31.76 ± 1.75) mg] and relative thyroid weight [(11.69 ± 3.47) mg/100 g] in DIT group was lower than that of the KIO3 group [(36.31 ± 5.23) mg,(12.83 ± 4.38) mg/100 g,all P ＜ 0.05].Conclusion Animal experimental results show that DIT has a better iodine-supplementing efficacy than that of KIO3.
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Classical swine fever (CSF) is a highly contagious disease among swine that has an important economic impact on worldwide. One clinical symptom of CSF is leukopenia, in particular lymphopenia, which is a characteristic event that occurs early in the course of CSF. Though lymphopenia associated with apoptosis, the pathogenic mechanism underlying the lymphopenia has not been well studied. To understand these mechanisms, we investigated the response of porcine B cell lines to infection with SW03, virulent strain isolated from swine tissue in Korea. This study demonstrated that SW03-infected L35 cell were induced apoptosis through the detection of activated caspase-3. In addition, SW03 infection leaded to alterations in pro-apoptotic, Bax, and anti-apoptotic, Bcl-xL proteins of Bcl-2 family. Our results would suggest that SW03-infected L35 cells induced apoptosis via intrinsic mitochondrial pathway.