顺铂对葡萄膜黑色素瘤细胞端粒酶活性的抑制

目的 研究顺铂对人葡萄膜黑色素瘤细胞端粒酶活性的抑制作用,为临床治疗人葡萄膜黑色素瘤提供理论依据.方法 采用特定时间下不同浓度以及特定浓度下不同作用时间的端粒酶抑制剂顺铂作用于体外培养的人黑色素瘤细胞.采用多聚酶链反应--酶联免疫吸附测定(PCR-ELISA)及聚丙烯酰胺凝胶电泳法(PCR-PAGE)测定细胞中端粒酶活性的变化.结果 作用72 h,端粒酶活性在顺铂浓度达到0.10mg/L后开始下降,当浓度达到1.00mg/L后其活性下降至(0.173±0.007).当顺铂浓度固定10.00 mg/L,顺铂作用时间达到24 h后开始出现抑制作用,48 h时达到抑制高峰,端粒酶活性下降至(0.276±0.024).随着顺铂浓度的增加及作用时间的延长,端粒酶活性逐渐下降(P＜0.05).PCR-PAGE显示顺铂浓度增加及作用时间延长,端粒酶活性的显色条带越来越少.结论 顺铂可有效降低体外培养的人眼葡萄膜黑色素瘤细胞端粒酶活性,并呈浓度和时间依赖性.     

IFN-α 2b对人脉络膜黑色素瘤细胞的毒性作用及对端粒酶活性的影响

目的：研究干扰素(IFN-α 2b)对人眼脉络膜黑色素瘤细胞端粒酶活性的影响及其对细胞的毒性作用,为临床化疗提供更多依据。方法:采用原代培养的人眼脉络膜黑色素瘤细胞做实验材料,用不同浓度IFN-α 2b、作用不同时间,设立对照组,用MTT法检测IFN-α 2b对细胞的毒性作用,PCR-ELISA法测定细胞中端粒酶活性的变化,并对两者关系进行相关分析。结果:随着IFN-α 2b浓度及作用时间的增加,细胞中端粒酶活性逐渐下降,在药物浓度大于5×104 IU/L及作用时间达到24 h后,其活性出现大幅度下降,随后出现细胞抑制率的明显升高,与端粒酶活性的下降具有一定的相关性。但均发生不同程度的滞后现象。结论:IFN-α 2b可作为一种端粒酶抑制剂,有效降低体外培养的人眼脉络膜黑色素瘤细胞的端粒酶活性,呈浓度和时间依赖性,并造成细胞死亡。     

端粒酶抑制剂对葡萄膜黑色素瘤细胞生长的抑制作用

Objective To study the restrain effects of two telomerase inhibitors included IFN-α2b (interferon-α2b)and CDDP(cisplatin;cis-dichlorodiamine platinum)to uveal melanoma cells of human eyes,and to provide theory bases for chemotherary of uveal melanoma.Methods Applied IFN-α2b and CDDP to the primary cultured uveal melanoma cells of human eyes individually with different concentration and duration,and compared with control group.Morphological changes were observed by inverted optical and electronic microscope,and the restrain effects were detected by MTT.Results More and more morphological changes of restraint and apoptosis of cells were observed with the increasing concentration and duration of IFN-α 2b and CDDP.When action durations were 72 h.the cells were inhibited with the concentration more than 500 IU/ml(IFN-α 2b)and 1.00 mg/L(CDDP)respectively.The cells restrain rates were(66.1±4.7)%and(74.0+3.3)%with the concentration more than 5000 IU/ml(IFN-α2b)and 10 mg/L (CDDP),and the rates were higher than before(P<0.001).When concentrations were 5000 IU/ml(IFN-α2b)and 10.00 mg/L(CDDP),cells were inhibited at 48 h and the restrain rates were(67.9±8.1)%and(76.5±1.61%at 72 h,respectively,(P相似文献   

端粒酶抑制剂对葡萄膜黑色素瘤细胞生长的抑制作用

Objective To study the restrain effects of two telomerase inhibitors included IFN-α2b (interferon-α2b)and CDDP(cisplatin;cis-dichlorodiamine platinum)to uveal melanoma cells of human eyes,and to provide theory bases for chemotherary of uveal melanoma.Methods Applied IFN-α2b and CDDP to the primary cultured uveal melanoma cells of human eyes individually with different concentration and duration,and compared with control group.Morphological changes were observed by inverted optical and electronic microscope,and the restrain effects were detected by MTT.Results More and more morphological changes of restraint and apoptosis of cells were observed with the increasing concentration and duration of IFN-α 2b and CDDP.When action durations were 72 h.the cells were inhibited with the concentration more than 500 IU/ml(IFN-α 2b)and 1.00 mg/L(CDDP)respectively.The cells restrain rates were(66.1±4.7)%and(74.0+3.3)%with the concentration more than 5000 IU/ml(IFN-α2b)and 10 mg/L (CDDP),and the rates were higher than before(P<0.001).When concentrations were 5000 IU/ml(IFN-α2b)and 10.00 mg/L(CDDP),cells were inhibited at 48 h and the restrain rates were(67.9±8.1)%and(76.5±1.61%at 72 h,respectively,(P相似文献   

端粒酶抑制剂对葡萄膜黑色素瘤细胞生长的抑制作用

Objective To study the restrain effects of two telomerase inhibitors included IFN-α2b (interferon-α2b)and CDDP(cisplatin;cis-dichlorodiamine platinum)to uveal melanoma cells of human eyes,and to provide theory bases for chemotherary of uveal melanoma.Methods Applied IFN-α2b and CDDP to the primary cultured uveal melanoma cells of human eyes individually with different concentration and duration,and compared with control group.Morphological changes were observed by inverted optical and electronic microscope,and the restrain effects were detected by MTT.Results More and more morphological changes of restraint and apoptosis of cells were observed with the increasing concentration and duration of IFN-α 2b and CDDP.When action durations were 72 h.the cells were inhibited with the concentration more than 500 IU/ml(IFN-α 2b)and 1.00 mg/L(CDDP)respectively.The cells restrain rates were(66.1±4.7)%and(74.0+3.3)%with the concentration more than 5000 IU/ml(IFN-α2b)and 10 mg/L (CDDP),and the rates were higher than before(P<0.001).When concentrations were 5000 IU/ml(IFN-α2b)and 10.00 mg/L(CDDP),cells were inhibited at 48 h and the restrain rates were(67.9±8.1)%and(76.5±1.61%at 72 h,respectively,(P相似文献   

端粒酶抑制剂对葡萄膜黑色素瘤细胞生长的抑制作用

Objective To study the restrain effects of two telomerase inhibitors included IFN-α2b (interferon-α2b)and CDDP(cisplatin;cis-dichlorodiamine platinum)to uveal melanoma cells of human eyes,and to provide theory bases for chemotherary of uveal melanoma.Methods Applied IFN-α2b and CDDP to the primary cultured uveal melanoma cells of human eyes individually with different concentration and duration,and compared with control group.Morphological changes were observed by inverted optical and electronic microscope,and the restrain effects were detected by MTT.Results More and more morphological changes of restraint and apoptosis of cells were observed with the increasing concentration and duration of IFN-α 2b and CDDP.When action durations were 72 h.the cells were inhibited with the concentration more than 500 IU/ml(IFN-α 2b)and 1.00 mg/L(CDDP)respectively.The cells restrain rates were(66.1±4.7)%and(74.0+3.3)%with the concentration more than 5000 IU/ml(IFN-α2b)and 10 mg/L (CDDP),and the rates were higher than before(P<0.001).When concentrations were 5000 IU/ml(IFN-α2b)and 10.00 mg/L(CDDP),cells were inhibited at 48 h and the restrain rates were(67.9±8.1)%and(76.5±1.61%at 72 h,respectively,(P相似文献   

端粒酶抑制剂对葡萄膜黑色素瘤细胞生长的抑制作用

Objective To study the restrain effects of two telomerase inhibitors included IFN-α2b (interferon-α2b)and CDDP(cisplatin;cis-dichlorodiamine platinum)to uveal melanoma cells of human eyes,and to provide theory bases for chemotherary of uveal melanoma.Methods Applied IFN-α2b and CDDP to the primary cultured uveal melanoma cells of human eyes individually with different concentration and duration,and compared with control group.Morphological changes were observed by inverted optical and electronic microscope,and the restrain effects were detected by MTT.Results More and more morphological changes of restraint and apoptosis of cells were observed with the increasing concentration and duration of IFN-α 2b and CDDP.When action durations were 72 h.the cells were inhibited with the concentration more than 500 IU/ml(IFN-α 2b)and 1.00 mg/L(CDDP)respectively.The cells restrain rates were(66.1±4.7)%and(74.0+3.3)%with the concentration more than 5000 IU/ml(IFN-α2b)and 10 mg/L (CDDP),and the rates were higher than before(P<0.001).When concentrations were 5000 IU/ml(IFN-α2b)and 10.00 mg/L(CDDP),cells were inhibited at 48 h and the restrain rates were(67.9±8.1)%and(76.5±1.61%at 72 h,respectively,(P相似文献   

端粒酶抑制剂对葡萄膜黑色素瘤细胞生长的抑制作用

Objective To study the restrain effects of two telomerase inhibitors included IFN-α2b (interferon-α2b)and CDDP(cisplatin;cis-dichlorodiamine platinum)to uveal melanoma cells of human eyes,and to provide theory bases for chemotherary of uveal melanoma.Methods Applied IFN-α2b and CDDP to the primary cultured uveal melanoma cells of human eyes individually with different concentration and duration,and compared with control group.Morphological changes were observed by inverted optical and electronic microscope,and the restrain effects were detected by MTT.Results More and more morphological changes of restraint and apoptosis of cells were observed with the increasing concentration and duration of IFN-α 2b and CDDP.When action durations were 72 h.the cells were inhibited with the concentration more than 500 IU/ml(IFN-α 2b)and 1.00 mg/L(CDDP)respectively.The cells restrain rates were(66.1±4.7)%and(74.0+3.3)%with the concentration more than 5000 IU/ml(IFN-α2b)and 10 mg/L (CDDP),and the rates were higher than before(P<0.001).When concentrations were 5000 IU/ml(IFN-α2b)and 10.00 mg/L(CDDP),cells were inhibited at 48 h and the restrain rates were(67.9±8.1)%and(76.5±1.61%at 72 h,respectively,(P相似文献   

端粒酶抑制剂对葡萄膜黑色素瘤细胞生长的抑制作用

Objective To study the restrain effects of two telomerase inhibitors included IFN-α2b (interferon-α2b)and CDDP(cisplatin;cis-dichlorodiamine platinum)to uveal melanoma cells of human eyes,and to provide theory bases for chemotherary of uveal melanoma.Methods Applied IFN-α2b and CDDP to the primary cultured uveal melanoma cells of human eyes individually with different concentration and duration,and compared with control group.Morphological changes were observed by inverted optical and electronic microscope,and the restrain effects were detected by MTT.Results More and more morphological changes of restraint and apoptosis of cells were observed with the increasing concentration and duration of IFN-α 2b and CDDP.When action durations were 72 h.the cells were inhibited with the concentration more than 500 IU/ml(IFN-α 2b)and 1.00 mg/L(CDDP)respectively.The cells restrain rates were(66.1±4.7)%and(74.0+3.3)%with the concentration more than 5000 IU/ml(IFN-α2b)and 10 mg/L (CDDP),and the rates were higher than before(P<0.001).When concentrations were 5000 IU/ml(IFN-α2b)and 10.00 mg/L(CDDP),cells were inhibited at 48 h and the restrain rates were(67.9±8.1)%and(76.5±1.61%at 72 h,respectively,(P相似文献   

端粒酶抑制剂对葡萄膜黑色素瘤细胞生长的抑制作用

Objective To study the restrain effects of two telomerase inhibitors included IFN-α2b (interferon-α2b)and CDDP(cisplatin;cis-dichlorodiamine platinum)to uveal melanoma cells of human eyes,and to provide theory bases for chemotherary of uveal melanoma.Methods Applied IFN-α2b and CDDP to the primary cultured uveal melanoma cells of human eyes individually with different concentration and duration,and compared with control group.Morphological changes were observed by inverted optical and electronic microscope,and the restrain effects were detected by MTT.Results More and more morphological changes of restraint and apoptosis of cells were observed with the increasing concentration and duration of IFN-α 2b and CDDP.When action durations were 72 h.the cells were inhibited with the concentration more than 500 IU/ml(IFN-α 2b)and 1.00 mg/L(CDDP)respectively.The cells restrain rates were(66.1±4.7)%and(74.0+3.3)%with the concentration more than 5000 IU/ml(IFN-α2b)and 10 mg/L (CDDP),and the rates were higher than before(P<0.001).When concentrations were 5000 IU/ml(IFN-α2b)and 10.00 mg/L(CDDP),cells were inhibited at 48 h and the restrain rates were(67.9±8.1)%and(76.5±1.61%at 72 h,respectively,(P相似文献   

不同期鼻咽癌病人血清中TNF－α和IFN－γ活性检测

本文研究不同期鼻咽癌病人血清中ＩＦＮ和ＴＮＦ的活性水平。结果如下：第Ⅰ、Ⅱ、Ⅲ、Ⅳ期各期病人，复发期病人和健康对照组的ＩＦＮ水平均值分别为５３．０，７１．４，６３．４，３２．８，５３．０和２１．０ＩＵ／ｍｌ，经方差分析各期病人和对照组之间有明显差异，Ｆ＝１１．６９９，Ｐ＜０．０００１。第Ⅰ－Ⅳ期各期病人以及复发期病人和健康对照组的ＴＮＦ水平均值分别为９９．５，１１５．３，１２１．２，２３５．４，８２．８和１１．４ＩＵ／ｍｌ。经方差分析各期病人和对照组之间有明显差异，Ｆ＝９．３９２８，Ｐ＜０．０１。值得注意的是第Ⅳ期病人的ＩＦＮ水平下降，而ＴＮＦ水平上升。中和试验确证病人血清中ＩＦＮ为γ型，ＴＮＦ为α型。本文讨论有关第Ⅳ期这两种细胞因子水平的分叉现象：ＩＦＮ－γ下降和ＴＮＦ－α上升与此肿瘤远距离转移和恶液质出现的关系。     

干扰素α对Jurkat细胞中CCR5表达的影响

目的:通过研究干扰素(IFN-α)对Jurkat E6-1细胞内CCR5 mRNA表达及基因表达调控的影响,探讨采用干扰素治疗相关疾病的方法。方法:用不同浓度的IFN-α刺激处于对数生长期的CD4+T淋巴细胞系Jurkat E6-1细胞。培养48小时后,提取细胞的总RNA,逆转录成cDNA,进行RT-PCR和Real time-PCR扩增目的基因CCR5;利用脂质体转染荧光素酶报告系统检测Jurkat E6-1细胞内CCR5活性变化情况。结果:(1)IFN-α在浓度为100 U/ml时对CCR5 mRNA的表达有明显抑制作用;在浓度为1 000 U/ml时对CCR5 mRNA的表达表现为增强作用;在浓度为10 000 U/ml时对CCR5 mRNA的表达增强作用有所减弱。(2)当IFN-α的浓度为100 U/ml时荧光素酶的活性最低;当IFN-α的浓度为1 000 U/ml时荧光素酶的活性最高;当IFN-α的浓度为10 000 U/ml时荧光素酶的活性又有所减低。结论:IFN-α作为一类很好的免疫调节、抗病毒生物制剂,不同剂量在一定范围内会对细胞CCR5的表达有显著影响。     

Tumor cell plasticity in uveal melanoma: microenvironment directed dampening of the invasive and metastatic genotype and phenotype accompanies the generation of vasculogenic mimicry patterns

The histological detection of laminin-rich vasculogenic mimicry patterns in human primary uveal melanomas is associated with death from metastases. We therefore hypothesized that highly invasive uveal melanoma cells forming vasculogenic mimicry patterns after exposure to a laminin-rich three-dimensional microenvironment would differentially express genes associated with invasive and metastatic behavior. However, we discovered that genes associated with differentiation (GDF15 and ATF3) and suppression of proliferation (CDKNa1/p21) were up-regulated in highly invasive uveal melanoma cells forming vasculogenic mimicry patterns, and genes associated with promotion of invasive and metastatic behavior such as CD44, CCNE2 (cyclin E2), THBS1 (thrombospondin 1), and CSPG2 (chondroitin sulfate proteoglycan; versican) were down-regulated. After forming vasculogenic mimicry patterns, uveal melanoma cells invaded only short distances, failed to replicate, and changed morphologically from the invasive epithelioid to the indolent spindle A phenotype. In human tissue samples, uveal melanoma cells within vasculogenic mimicry patterns assumed the spindle A morphology, and the expression of Ki67 was significantly reduced in adjacent melanoma cells. Thus, the generation of vasculogenic mimicry patterns is accompanied by dampening of the invasive and metastatic uveal melanoma genotype and phenotype and underscores the plasticity of these cells in response to cues from the microenvironment.     

干扰素-γ对乳腺癌细胞HLA-Ⅰ分子表达的影响

目的 研究干扰素-γ（IFN-γ）对乳腺癌细胞表面HLA-Ⅰ类分子表达的影响,并探讨其在乳腺癌治疗中的免疫作用机制。方法 培养乳腺癌SKBR3细胞株,根据IFN-γ浓度将其分为25 U/ml、50 U/ml、100 U/ml组,未加药组为空白对照组。通过实时荧光定量PCR和流式细胞术实验检测经不同浓度IFN-γ作用后的乳腺癌SKBR3细胞内HLA-Ⅰ mRNA的含量,以及表面的HLA-Ⅰ类抗原的表达。结果 实时荧光定量PCR结果显示,空白对照组、25 U/ml、50 U/ml、100 U/ml组HLA-Ⅰ mRNA的表达量分别为（1.00±0.00）、（2.26±0.54）、（3.89±0.81）、（5.94±0.97）,三个浓度组均较空白对照组表达量高（P＜0.05）;流式细胞术结果显示,空白对照组、25 U/ml、50 U/ml、100 U/ml组细胞表面HLA-Ⅰ蛋白表达量为（14.87±2.80）、（44.33±3.36）、（48.80±2.63）、（56.77±1.93）,三个浓度组均高于对照组（P＜0.05）。结论 IFN-γ上调乳腺癌细胞HLA-Ⅰ类抗原的表达有利于抗乳腺癌作用。     

IL-23在慢性乙型肝炎免疫调节中的作用研究

目的 探讨IL-23在慢性乙型肝炎免疫调节中的作用。方法 选取我院收治的32例HBeAg阳性慢性乙型肝炎患者为研究对象,根据ALT水平分为ALT≥120 IU/ml患者16例,ALT＜120 IU/ml患者16例,另外选择我院健康体检中心20例体检者作为健康对照组,采用酶联免疫分析法（ELISA）检测血清IL-23水平,采用流式细胞仪检测Th17细胞百分率。结果 与健康对照组相比,HBeAg阳性慢性乙型肝炎患者血清IL-23表达及外周血Th17细胞百分率均升高,差异有统计学意义（P＜0.05）; ALT≥120 IU/ml患者血清IL-23浓度（394.81±101.84）pg/ml高于ALT＜120IU/ml的（283.69±85.65）pg/ml,ALT≥120 IU/ml患者Th17细胞百分率（3.25±0.70）%高于ALT＜120IU/ml的（2.68±0.61）%,差异均有统计学意义（P＜0.05）;慢性乙型肝炎患者外周血Th17细胞百分率、血清IL-23浓度与ALT程度呈正相关（P均＜0.05）;Th17细胞百分率与血清IL-23浓度呈正相关（P＜0.05）。结论 IL-23可能通过影响Th17细胞的免疫调节参与慢性乙型肝炎患者炎症,为临床提供了新的靶标。     

恶性淋巴瘤患者TH 1/TH 2细胞因子表达水平的研究

目的 探讨恶性淋巴瘤患者血清中TH1/TH2细胞因子变化及其临床意义,为肿瘤的免疫治疗提供实验依据.方法 用流式细胞小球微阵列术(cytometric bead array,CBA)检测92例恶性淋巴瘤患者及70例健康人群血清中γ干扰素(IFN-γ)、肿瘤坏死因子-α仪(TNF-α)、白细胞介素(IL-2、IL-4、IL-5、IL-10)表达水平.结果 92例恶性淋巴瘤患者血清中TH1型细胞因子的水平分别为:IFN-γ(34.26±33.4g)pg/ml、TNF-α(8.17±10.09)pg/ml、IL-2(3.74 4±1.72)pg/ml;TH2型细胞因子的水平分别为:IL-10(6.28±8.56)pg/ml、IL-5(3.53±3.20)pg/ml、IL-4(6.22±7.13)pg/ml.除TNF-α表达水平降低外,其余5项均明显高于健康体检组,差异有统计学意义(P＜0.01).TH1细胞因子IL-2与TH2细胞因子IL-4的比值明显下降(0.78±O.44),与健康体检组(1.09±0.45)比较差异有统计学意义(P＜0.01).IL-10与疾病的进展相关,Ⅲ/Ⅳ期恶性淋巴瘤患者的表达水平为(9.58±13.96)pg/ml,Ⅰ/Ⅱ期的表达水平为(4.77±3.50)pg/ml,二者比较差异有统计学意义(P＜0.01).IFN-γ在大于60岁的恶性淋巴瘤患者中表达水平明显降低,与其他年龄段恶性淋巴瘤患者比较差异有统计学意义(P ＜0.05).结论 恶性淋巴瘤患者血清中TH1/TH2细胞因子平衡失调,检测TH1/TH2细胞因子可作为评价淋巴瘤临床进展及预后指标.TH1/TH2平衡向TH2方向漂移,这可能是肿瘤细胞发生免疫逃逸,从而导致肿瘤的发生或者转移的原因之一.     

Abrasive Endoprosthetic Wear Particles Inhibit IFN-γ Secretion in Human Monocytes Via Upregulating TNF-α-Induced miR-29b

The adverse biological responses to prostheses wear particles commonly led to the failure of total hip arthroplasty. Among the released cytokines, interferon-γ (IFN-γ) has been found to be a critical functional factor during osteoclast differentiation. However, the molecular mechanism underlying the regulation of IFN-γ in wear particles-induced cells still needs to be determined. Four kinds of abrasive endoprosthetic wear particle were used to treat THP-1 cells, including polymethylmethacrylate (PMMA), zirconiumoxide (ZrO₂), commercially pure titanium (cpTi), and titanium alloy (Ti-6Al-7Nb), with a concentration of 0.01, 0.05, 0.1, or 0.2 mg/ml for 48 h. The expression of IFN-γ and miR-29b was detected by real-time RT-PCR or ELISA. Luciferase reporter assay was performed to determine the regulation of miR-29b on IFN-γ. The effect of miR-29b inhibitor on the expression of wear particle-induced IFN-γ was detected. The expression of miR-29b was examined in THP-1 cells treated with tumor necrosis factor-alpha (TNF-α). The expression of IFN-γ was downregulated and the level of miR-29b was increased in THP-1 cells pretreated with wear particles. IFN-γ was a target of miR-29b. Wear particles inhibited the expression of IFN-γ through miR-29b. The expression of miR-29b was significantly reduced in THP-1 cells treated with TNF-α neutralizing antibody and particles comparing to that in the cells treated with particles alone. Wear particles inhibit the IFN-γ secretion in human monocytes, which was associated with the upregulating TNF-α-induced miR-29b.