1.Regulatory role and mechanism of miR-383 in bortezomib-mediated in vitro inhibition of osteosarcoma
Kaizhong HU ; Shaozhi ZHENG ; Fenting JIA ; Chuanyi BAI ; Li ZHANG
Journal of Chinese Physician 2025;27(5):693-698
Objective:To investigate the expression of miR-383(Micro RNA-383)in osteosarcoma cells and to verify whether upregulation of miR-383 can enhance the therapeutic efficacy of bortezomib against osteosarcoma.Methods:Fluorescence in situ hybridization (FISH) was used to detect the expression of miR-383 in osteosarcoma and normal bone tissues. Real-time quantitative polymerase chain reaction (qRT-PCR) was employed to measure the expression of miR-383 in different osteosarcoma cell lines (SaoS-2, HOS, U-2OS, and MG63)and the osteoblast cell line hFOB 1.19.The proliferative capacity of osteosarcoma cells treated with 5 nmol/L and 10 nmol/L bortezomib was assessed using the cell counting kit-8 (CCK-8) with dimethyl sulfoxide (DMSO) as a control. The activity of caspase-3 was also measured. HOS and MG63 cells were treated with DMSO, bortezomib, miR-383 mimics, or negative controls, and the proliferative capacity and apoptosis levels were re-evaluated using CCK-8 and flow cytometry, respectively.Results:FISH results showed that the level of miR-383-5p in osteosarcoma tissues was significantly lower than that in normal bone tissues ( P<0.05). qRT-PCR results indicated that miR-383 levels in osteosarcoma cells (MG63, HOS, Saos-2, U-2OS) were lower than those in osteoblasts (hFOB1.19), with significant differences among different osteosarcoma cell lines(all P<0.05).The lowest levels of miR-383 were observed in HOS and MG63 cells. CCK-8 and caspase-3 activity assays revealed that among the cells treated with DMSO and two doses of bortezomib, HOS and MG63 cells had higher baseline proliferative capacity. Compared with DMSO-treated control cells, cells treated with 5 nmol/L and 10 nmol/L bortezomib exhibited inhibited proliferation (all P<0.05) and increased caspase-3 activity (all P<0.05). The effect of 10 nmol/L bortezomib was stronger than that of 5 nmol/L (all P<0.05). Compared with negative control-transfected cells, osteosarcoma cells (MG63 and HOS) with overexpressed miR-383 showed inhibited proliferation and increased apoptosis levels (all P<0.05). After bortezomib treatment, osteosarcoma cells (MG63 and HOS)with overexpressed miR-383 exhibited reduced proliferative capacity and enhanced apoptosis levels (all P<0.05). Conclusions:miR-383 exerts anticancer effects in osteosarcoma by inhibiting cell proliferation. Its overexpression significantly enhances the therapeutic efficacy of bortezomib, offering a new direction for the treatment strategies of osteosarcoma.
2.Regulatory role and mechanism of miR-383 in bortezomib-mediated in vitro inhibition of osteosarcoma
Kaizhong HU ; Shaozhi ZHENG ; Fenting JIA ; Chuanyi BAI ; Li ZHANG
Journal of Chinese Physician 2025;27(5):693-698
Objective:To investigate the expression of miR-383(Micro RNA-383)in osteosarcoma cells and to verify whether upregulation of miR-383 can enhance the therapeutic efficacy of bortezomib against osteosarcoma.Methods:Fluorescence in situ hybridization (FISH) was used to detect the expression of miR-383 in osteosarcoma and normal bone tissues. Real-time quantitative polymerase chain reaction (qRT-PCR) was employed to measure the expression of miR-383 in different osteosarcoma cell lines (SaoS-2, HOS, U-2OS, and MG63)and the osteoblast cell line hFOB 1.19.The proliferative capacity of osteosarcoma cells treated with 5 nmol/L and 10 nmol/L bortezomib was assessed using the cell counting kit-8 (CCK-8) with dimethyl sulfoxide (DMSO) as a control. The activity of caspase-3 was also measured. HOS and MG63 cells were treated with DMSO, bortezomib, miR-383 mimics, or negative controls, and the proliferative capacity and apoptosis levels were re-evaluated using CCK-8 and flow cytometry, respectively.Results:FISH results showed that the level of miR-383-5p in osteosarcoma tissues was significantly lower than that in normal bone tissues ( P<0.05). qRT-PCR results indicated that miR-383 levels in osteosarcoma cells (MG63, HOS, Saos-2, U-2OS) were lower than those in osteoblasts (hFOB1.19), with significant differences among different osteosarcoma cell lines(all P<0.05).The lowest levels of miR-383 were observed in HOS and MG63 cells. CCK-8 and caspase-3 activity assays revealed that among the cells treated with DMSO and two doses of bortezomib, HOS and MG63 cells had higher baseline proliferative capacity. Compared with DMSO-treated control cells, cells treated with 5 nmol/L and 10 nmol/L bortezomib exhibited inhibited proliferation (all P<0.05) and increased caspase-3 activity (all P<0.05). The effect of 10 nmol/L bortezomib was stronger than that of 5 nmol/L (all P<0.05). Compared with negative control-transfected cells, osteosarcoma cells (MG63 and HOS) with overexpressed miR-383 showed inhibited proliferation and increased apoptosis levels (all P<0.05). After bortezomib treatment, osteosarcoma cells (MG63 and HOS)with overexpressed miR-383 exhibited reduced proliferative capacity and enhanced apoptosis levels (all P<0.05). Conclusions:miR-383 exerts anticancer effects in osteosarcoma by inhibiting cell proliferation. Its overexpression significantly enhances the therapeutic efficacy of bortezomib, offering a new direction for the treatment strategies of osteosarcoma.
3.Clinical and epidemic characteristics of 27 cases with Brucellosis confirmed by bacterial culture
Kaizhong LUO ; Wei CAO ; Min WANG ; Shixiong HU ; Shaojun ZHANG ; Guozhong GONG
Journal of Chinese Physician 2017;19(11):1654-1657
Objective To improve the diagnostic and therapeutic ability of Brucellosis by analyzing the epidemic and clinical characteristics.Methods A retrospective analysis was done on the data of Brucellosis patients treated in our hospital from 2007 to 2016.Results Since the first case was diagnosed in 2012,27 patients [19 male and 8 female,mean age (44.4 ± 16.9) years] were confirmed by clinical manifestations and positive bacterial cultures results.The annual number of cases from 2012 to 2016 was 1,1,6,4 and 15.Among them,10 cases (37.0%) had a history of close contact with goat,7 cases (25.9%)with raw mutton,1 case (3.7%) with raw beef and 1 case (3.7%) with suspicious laboratory contamination while 8 cases (29.6%) had no evident risk factors for Brucellosis.The common clinical manifestation included fever (81.5%),lumbago/joint pain (55.6%),fatigue (33.3%) and hyperhidrosis (22.2%).The white blood cell count was normal among 20 cases (74.1%) while 6 cases (22.2%) with leukopenia.Mild to moderate anemia in 20 cases (74.1%) and decreased platelet number in 4 case (14.8%).The percentage of elevated alanine aminotransferase,aspartate aminotransferase and lactate dehydrogenase was 32.0%,48.0% and 100%,respectively.Decreased albumin level was found in 23 cases (92.0%).The percentage of elevated erythrocyte sedimentation rate (ESR),C-reactive protein and serum ferritin 75.0%,82.3% and 77.8%,respectively,while 12 cases (85.7%) with procalcitonin level below 0.5 ng/L.According to follow-up for at least half year,all the cases were cured by active medical management.Conclusions The number of Brucellosis cases is rapidly increasing in our hospital.It's of great significance to know the epidemic and clinical characteristics of Brucellosis.

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