结缔组织生长因子促进人卵巢癌细胞系SKOV3增殖与迁移

目的 探讨结缔组织生长因子(CTGF)对人卵巢癌细胞系SKOV3增殖、迁移能力的影响,并探索其可能机制.方法 用重组人CTGF(rhCTGF)处理SKOV3细胞;利用CCK-8法和集落形成实验检测SKOV3细胞的增殖;利用细胞划痕实验检测SKOV3细胞的迁移;Western blot检测上皮-间充质转化(EMT)相关蛋...     

NEDD8 共价修饰抑制因子MLN4924 对人卵巢癌细胞株SKOV3 增殖和凋亡的影响

目的:探讨神经前体细胞发育相关受控表达分子8(NEDD8)共价修饰抑制因子MLN4924 对人卵巢癌细胞增殖和凋亡的影响,并分析其可能机制。方法:使用不同浓度MLN4924(0、0.125、0.25、0.5 μmol/ L) 处理人卵巢癌细胞株SKOV3 4 h,免疫印迹法检测PAR3、HER2、Neddylated-cullins、P鄄I资B琢的表达水平;同时取细胞培养上清用ELISA 检测IL-6 的分泌。不同浓度MLN4924 处理SKOV3 细胞72 h,CCK8 法检测细胞增殖抑制率,流式细胞仪检测细胞周期和细胞凋亡率。结果:MLN4924 作用4 h 时,能够使Neddylated-cullins 下降、P-κBα积聚,IL-6 分泌减少,而PAR3、HER2 表达水平变化不显著。MLN4924 在0.125、0.25 和0.5 μmol/ L 的浓度下处理SKOV3 细胞72 h 时,对细胞增殖有明显的抑制作用,并且随着剂量的加大,抑制效果也显著提高;同时加药组S 期细胞百分率相较于对照组明显升高,且形成大量四倍体,使之没有完整的细胞周期;细胞凋亡率也随MLN4924 剂量增大而升高。结论:NEDD8 修饰特异性抑制剂MLN4924 能够明显抑制人卵巢癌细胞株SKOV3 的增殖,诱导细胞周期阻滞和凋亡。上述效应可能与NF-κB 通路活性受到抑制和IL-6 表达下降有关。     

重组腺病毒Ad-IL-12感染淋巴细胞对人卵巢癌细胞SKOV3增殖和凋亡的影响

目的 探讨重组腺病毒Ad-IL-12感染人外周血淋巴细胞后对人卵巢癌细胞SKOV3周期、增殖和凋亡的影响.方法 采用重组人IL-12腺病毒(Ad-IL-12)感染人外周血淋巴细胞,感染48 h后与人卵巢癌细胞SKOV3共培养(SKOV3/Ad-IL-12),同时设未感染组(SKOV3)和Ad-GFP空载感染组(SKOV3/Ad-GFP)作为对照.RT-PCR检测IL-12双亚基P35和P40的mRNA表达,ELISA检测细胞培养上清中IL-12 P70蛋白的分泌,CCK8法检测卵巢癌细胞增殖,流式细胞术分析卵巢癌细胞周期和凋亡,RT-PCR和Western blot检测抗凋亡蛋白BCL-2、survivin 的表达.结果 重组腺病毒Ad-IL-12可成功感染人外周血淋巴细胞,分泌较高水平的IL-12 P70蛋白;淋巴细胞过表达IL-12可抑制SKOV3细胞的增殖;与未感染组和空载组相比,SKOV3/Ad-IL-12组G1期细胞显著增加,凋亡率显著升高(P＜0.05),SKOV3细胞中抗凋亡蛋白BCL-2和survivin显著下调(P＜0.05).结论 人IL-12基因重组腺病毒可以成功感染人外周血淋巴细胞,分泌IL-12 P70蛋白,并可以通过阻滞细胞周期进程和促进细胞凋亡抑制人卵巢癌细胞SKOV3的增殖.     

青蒿素对人膀胱癌T24细胞增殖、凋亡、迁移与侵袭的影响

目的探讨青蒿素对人膀胱癌T24细胞增殖、凋亡、迁移与侵袭的影响及可能机制。方法将人膀胱癌T24细胞分为空白对照组与青蒿素组(50、100、200μmol/L)。不同浓度青蒿素干预48h后,MTT方法检测T24细胞的增殖活力,流式细胞术检测T24细胞的凋亡率。200μmol/L青蒿素干预48 h后,划痕实验与Transwell侵袭实验检测青蒿素对T24细胞迁移与侵袭能力的影响;Western blot实验检测T24细胞中PI3K、p-Akt与pmTOR的表达。结果不同浓度青蒿素均能显著抑制T24细胞增殖,并促进细胞凋亡,且呈浓度依赖性(P0.01)。200μmol/L青蒿素干预48 h后,T24细胞迁移与侵袭能力显著降低,T24细胞中PI3K的表达以及Akt和mTOR的磷酸化水平降低(P0.01)。结论青蒿素能够通过抑制PI3K/Akt/mTOR信号通路抑制膀胱癌T24细胞增殖、迁移与侵袭,并促进凋亡。     

LAIR-1对人急性髓系白血病细胞HEL增殖、凋亡和迁移的影响

研究人白细胞相关免疫球蛋白样受体1(leukocyte-associated immunoglobulin-like receptor 1,LAIR-1)对人红白血病细胞HEL增殖、凋亡和迁移的影响,探讨LAIR-1在急性髓系白血病(acute myeloid leukemia,AML)中的作用。采用LAIR-1慢病毒表达载体(LV-LAIR-1)转染HEL细胞,建立稳定高表达LAIR-1的HEL细胞株,分别采用流式细胞术、激光扫描共聚焦显微技术、western blotting和RT-PCR鉴定LAIR-1分子的表达效率。采用CCK-8试剂盒、FITC Annexin V凋亡试剂盒和Transwell迁移实验分别检测LAIR-1对HEL细胞增殖、凋亡和迁移能力的影响。实验结果显示,经LVLAIR-1转染的HEL细胞高表达LAIR-1分子,阳性率达到85%以上;功能实验结果显示,与对照组细胞相比,LAIR-1能够显著抑制HEL细胞的增殖(P<0.001),并且促进其凋亡(P<0.01),但是对细胞迁移能力无明显影响(P>0.05)。上述研究结果显示,LAIR-1分子的表达能够影响人AML细胞的生物学功能。     

ELF脉冲电磁场对脑组织钙离子迁移量的影响

目的研究极低频(extremely low frequency,ELF)脉冲电磁场对脑组织钙离子迁移量的影响。方法以鸡脑组织为生物学实验对象,以同位素示踪法为标记方法,用液体闪烁计数器为检测仪器,实验观察ELF脉冲电磁场对脑组织钙离子迁移量的影响。结果不同特征外加ELF脉冲电磁场对脑组织钙离子迁移量影响不同,如频率为16 Hz、场强为42.5 V/m的ELF脉冲电磁场可引起脑组织钙离子迁移量显著变化。频率为16 Hz、场强为62.2 V/m与频率为31 Hz、场强为42.5 V/m的ELF脉冲电磁场却未引起脑组织钙离子迁移量显著改变。结论只有频率和强度恰当组合的ELF电磁场才可引起脑组织钙离子迁移量显著升高,呈现出生物学窗效应。另一方面在既注重物理学分析又注重生物系统特性基础上,从膜通道内的带电粒子动力学方程入手分析讨论生物学窗效应产生机制。     

紫草素对人膀胱癌T24细胞增殖、凋亡、迁移与侵袭的影响

目的探讨紫草素对人膀胱癌T24细胞增殖、凋亡、迁移与侵袭的影响及可能机制。方法将人膀胱癌T24细胞分为空白对照组与紫草素组(10、20、40μmol/L)。不同浓度紫草素干预48h后,MTT方法检测T24细胞的增殖活力,流式细胞术检测T24细胞的凋亡率。40μmol/L紫草素干预48 h后,Transwell实验检测紫草素对T24细胞迁移与侵袭能力的影响;Western blot实验检测T24细胞中半胱氨酸天冬氨酸蛋白水解酶3/9(Caspase-3/Caspase-9)与基质金属蛋白酶2/9(MMP-2/MMP-9)的表达。结果不同浓度紫草素均能显著抑制T24细胞增殖,并促进细胞凋亡,且呈浓度依赖性(P0.01)。40μmol/L紫草素干预48 h后,T24细胞迁移与侵袭能力显著降低,T24细胞中Caspase-3、Caspase-9、MMP-2与MMP-9的蛋白表达水平均显著升高(P0.01)。结论紫草素能够抑制膀胱癌T24细胞增殖、迁移与侵袭,并促进凋亡。     

丹参二萜醌通过调控CCL3影响骨髓瘤细胞的增殖、凋亡和迁移

目的 探讨丹参二萜醌对骨髓瘤细胞的增殖、凋亡和迁移的影响和分子机制。方法 采用不同浓度的丹参二萜醌体外处理骨髓瘤细胞RPMI8226。四甲基偶氮唑蓝(MTT)实验检测细胞增殖活力;集落形成实验检测细胞克隆形成能力;流式细胞术检测细胞凋亡;Transwell实验检测细胞迁移能力;Western blot检测P21、半胱氨酸天冬氨酸蛋白酶(Caspase-3)、上皮细胞钙粘蛋白(E-cadherin)和趋化因子配体3(CCL3)蛋白的表达;实时荧光定量PCR(RT-qPCR)检测CCL3 mRNA的表达。将CCL3小干扰RNA(si-CCL3)转染RPMI8226,采用上述方法检测细胞增殖、克隆形成、凋亡、迁移能力变化。结果 丹参二萜醌处理后,RPMI8226细胞存活率、克隆形成数目、细胞迁移数目显著降低,细胞凋亡率显著升高,P21、Caspase-3和E-cadherin蛋白的表达显著升高,并呈明显的剂量依赖关系(P<0.05)。转染si-CCL3后,RPMI8226细胞存活率、克隆形成数目、细胞迁移数目显著降低,细胞凋亡率显著升高,P21、Caspase-3和E-cadherin...     

黄连素抑制人卵巢癌SKOV3细胞增殖并诱导凋亡

黄连素(berberine)对多种肿瘤有抑制增殖和诱导凋亡的作用[1-2],但在卵巢癌中的作用尚未见报道.本研究以人卵巢癌SKOV3细胞为研究对象,观察黄连素对SKOV3细胞增殖抑制作用及凋亡诱导效应.     

3-溴丙酮酸对人卵巢癌细胞增殖和凋亡的影响

目的 观察3-溴丙酮酸对人卵巢癌SK-OV-3细胞株增殖和凋亡的影响,探讨3-溴丙酮酸对人卵巢癌细胞的体外抗肿瘤效应.方法 MTT比色法和集落形成试验检测3-溴丙酮酸对SK-OV-3细胞的增殖抑制作用;采用投射电镜观察3-溴丙酮酸作用后的SK-OV-3细胞形态学改变;流式细胞术检测3-溴丙酮酸作用后SK-OV-3细胞的凋亡,并对细胞周期的变化进行分析.结果 3-溴丙酮酸对SK-OV-3细胞的增殖有明显的抑制作用,具有时间（一段时间内）和剂量依赖性（P＜0.05）.3-溴丙酮酸主要将细胞阻滞于G0/G1,S期细胞明显减少.结论 3-溴丙酮酸对SK-OV-3细胞增殖的抑制效应具有时间依赖性（一段时间内）和剂量依赖性,并可诱导其凋亡.     

FTO accelerates ovarian cancer cell growth by promoting proliferation,inhibiting apoptosis,and activating autophagy

ObjectiveFat mass and obesity-associated protein (FTO) is identified as a critical demethylase involved in various physiological processes. Despite efforts have been made to study the biological functions of FTO in certain cancers, the role of FTO in ovarian cancer is largely unknown. In this study, we sought to investigate the function of FTO on proliferation, apoptosis and autophagy of ovarian cancer cells.MethodsQuantitative real-time PCR was performed to detect FTO expression in ovarian tumor tissues and ovarian cancer cell lines OVCAR-3, SKOV-3, COC1, HO-8910 and A2780. SKOV-3 cells were constructed with FTO overexpression and A2780 cells were constructed with FTO knockdown. CCK-8 assay was used to examine cell viability and flow cytometry was used to detect cell apoptosis. Activity assay kits were applied to detect caspase-3 and caspase-9 levels. Western blot was performed to measure the expressions of FTO, PCNA, Bax, Bcl-2, LC3, ATG5, P62, p-AKT and AKT. Stable FTO-overexpression SKOV-3 cells or FTO-depletion A2780 cells were injected subcutaneously into male Balb/c-nu mice. Xenografted tumors were assayed by H&E staining. Immunohistochemistry was subjected to measure FTO and Ki67 expressions.ResultsFTO was up-regulated in ovarian tumor tissues compared with non-cancerous ovarian tissues. FTO overexpression markedly increased viability and autophagy function, but decreased apoptosis of ovarian cancer cells. In addition, FTO overexpression promoted AKT phosphorylation. In contrast, FTO silence showed the opposite effect.ConclusionFTO accelerated ovarian cancer cell growth by promoting proliferation, inhibiting apoptosis, and activating autophagy.     

The effects of pulsed electromagnetic field on the functions of osteoblasts on implant surfaces with different topographies

The use of pulsed electromagnetic fields (PEMFs) is a promising approach to promote osteogenesis. However, few studies have reported the effects of this technique on the osseointegration of endosseous implants, especially with regard to different implant topographies. We focused on how the initial interaction between cells and the titanium surface is enhanced by a PEMF and the possible regulatory mechanisms in this study. Rat osteoblasts were cultured on three types of titanium surfaces (Flat, Micro and Nano) under PEMF stimulation or control conditions. Protein adsorption was significantly increased by the PEMF. The number of osteoblasts attached to the surfaces in the PEMF group was substantially greater than that in the control group after 1.5 h incubation. PEMF stimulation oriented the osteoblasts perpendicular to the electromagnetic field lines and increased the number of microfilaments and pseudopodia formed by the osteoblasts. The cell proliferation on the implant surfaces was significantly promoted by the PEMF. Significantly increased extracellular matrix mineralization nodules were observed under PEMF stimulation. The expression of osteogenesis-related genes, including BMP-2, OCN, Col-1,ALP, Runx2 and OSX, were up-regulated on all the surfaces by PEMF stimulation. Our findings suggest that PEMFs enhance the osteoblast compatibility on titanium surfaces but to different extents with regard to implant surface topographies. The use of PEMFs might be a potential adjuvant treatment for improving the osseointegration process.     

槲皮素对人卵巢癌SKOV-3细胞增殖的影响

 目的：探讨槲皮素对人卵巢癌SKOV-3细胞增殖抑制和凋亡的影响,为卵巢癌临床治疗提供依据。方法：不同浓度槲皮素处理卵巢癌SKOV-3细胞后,采用MTT实验检测细胞增殖抑制作用并计算抑制率,细胞免疫化学染色法鉴定细胞凋亡,流式细胞术检测细胞周期及细胞凋亡。结果：槲皮素能够抑制SKOV-3细胞的增殖,且呈时间和剂量依赖性,免疫荧光显示槲皮素对SKOV-3细胞具有诱导凋亡作用,流式细胞术显示SKOV-3细胞被阻滞在S期, G2/M期细胞比例降低,凋亡率上升。结论：槲皮素在体外能够抑制卵巢癌SKOV-3细胞的增殖,阻止细胞由S期向G2期移行,促进其凋亡。     

Paeonol induces apoptosis in human ovarian cancer cells

Paeonol is a broad-spectrum antitumor agent, which is widely used in the treatment of various tumors in Asia. However, the effect of paeonol on ovarian cancer remains unclear. The purpose of this study was to investigate the effect of paeonol on ovarian cancer cells and its possible mechanism. Results measured by MTT (methyl thiazoyltetrazolium) assay showed that cell viability was markedly reduced in a dosage-dependent manner, when treated with paeonol for 24 h. Flow cytometry and Hoechst staining results indicated that the rate of apoptosis in the paeonol pretreatment group was higher than the control group. After co-culture with paeonol, cleaved Caspase 3 protein levels increased while survivin protein levels decreased. In conclusion, our findings indicate that paeonol can induce apoptosis of ovarian cancer cells via activation of Caspase 3 and down-regulation of survivin, and therefore is potentially an effective chemotherapeutic agent for ovarian cancer.     

The expression of Bcl-2 in adenomyosis and its effect on proliferation,migration, and apoptosis of endometrial stromal cells

Adenomyosis is a common gynecologic disease that severe impact on women. Previous studies have found that Bcl-2 abnormally expressed in adenomyosis. However, the exact mechanisms of Bcl-2 in the pathogenesis of adenomyosis are unclear. In this study, we are to explore the effect of Bcl-2 on proliferation, migration, and apoptosis of endometrial stromal cells. The expression of Bcl-2 were evaluated by Western blot and RT-qPCR. We used RNA interference to silence Bcl-2 gene of endometrial stromal cells, and then Cell Counting Kit(CCK-8), cell scratch repair test, and Annexin V-APC/propidium iodide (PI) staining were performed to detect the cell viability, migration ability and apoptotic rate. The results of the present study revealed that the expression of Bcl-2 was evidently higher than that in control group. After silencing the Bcl-2 gene, the cytoactive and migration ability of endometrial stromal cells of adenomyosis decreased, and the apoptotic rate increased. In conclusion, Bcl-2 is overexpressed in adenomyosis and participate in the pathogenesis of adenomyosis. Bcl-2 may be a potential novel target for adenomyosis treatment.     

丙泊酚对人胶质瘤U251细胞株增殖、凋亡、侵袭和转移能力的调节作用

目的　探究丙泊酚对人胶质瘤U251细胞增殖、凋亡、侵袭和转移能力的作用和机制。 方法 将细胞随机分为U251组、Propofol（1 mM）组、Propofol（2 mM）组和Propofol（5 mM）组。除U251组外,其余组用相应浓度的丙泊酚处理,CCK8检测细胞增殖,流式检测细胞凋亡,Transwell检测细胞侵袭能力,划痕实验检测细胞迁移能力,Western blot检测Ki67、Caspase-3、血管内皮生长因子（Vascular endothelial growth factor, VEGF）、磷脂酰肌醇-3-羟激酶（Phosphoinositide 3-kinase, PI3K）、Akt、p-PI3K和p-Akt的表达。 结果 与U251组比较,Propofol （1, 2, 5 mM）组细胞增殖速度明显降低,细胞凋亡率显著升高;同时,Propofol （1, 2, 5 mM） 组侵袭细胞数与U251组比较明显减少,Propofol（2, 5 mM） 组划痕闭合率明显低于U251组;此外,丙泊酚还能显著抑制细胞增殖和迁移相关蛋白Ki67和VEGF表达,诱导细胞凋亡相关蛋白Caspase-3表达;丙泊酚能明显降低p-PI3K/PI3K和p-Akt/Akt的比值。 结论 丙泊酚能降低胶质瘤U251细胞的增殖、侵袭和转移能力,抑制U251细胞凋亡,作用机制可能与抑制PI3K/Akt信号通路激活有关。     

脉冲电磁场辅助骨细胞培养装置的研制

目的 根据脉冲电磁场对骨细胞的作用机制,设计了能够产生均匀磁场、参数可调的脉冲电磁场发生装置和磁场作用板;叙述了实验装置的硬件构成、软件控制功能以及磁场作用板的设计,并通过试验测试,观察记录了在未加入磁性材料和加入磁性材料后磁场的分布情况.方法 设计了强度为4～6 mT、频率8 Hz的参数可调的脉冲电磁场发生装置和专用的适合细胞培养的磁场作用装置,并对产生的磁场进行了实验测定.结果 实验装置运行时,对各参数检测表明,实验装置满足设计的要求:在加入磁性材料后,磁场作用表面的磁场分布均匀.结论 实验装置满足设计要求,可为进一步进行骨细胞培养提供可靠的实验手段.