白藜芦醇诱导人成骨肉瘤细胞凋亡及其机制

目的 探讨白藜芦醇(resveratrol)诱导人成骨肉瘤细胞株MG63的凋亡作用及其分子机制.方法 应用不同浓度的白藜芦醇作用于MG63细胞,利用透射电镜观察细胞形态学变化;采用四甲基偶氮唑蓝(MTT)法检测药物对细胞的增殖抑制率;流式细胞术(FCM)检测细胞凋亡、细胞周期分布;反转录聚合酶链反应(RT-PCR)检测P21cip1/WAF1 mRNA、Survivin mRNA的表达.结果 白藜芦醇能明显抑制MG63细胞的增殖,并呈剂量和时间依赖性.经白藜芦醇处理MG 63细胞后,透射电镜下可见到凋亡细胞,FCM分析发现,10、20 μg/ml的白藜芦醇处理细胞72 h后,凋亡率分别为(11.9±0.6)%、(19.7%±0.9)%(P＜0.01),处理后的细胞G0/G1期比例增高,S期、G2/M期细胞比例减少;RT-PCR检测到P21cip1/WAF1表达升高,Survivin表达降低(P＜0.01).结论 白藜芦醇能明显抑制MG63细胞的增殖,诱导其凋亡,其机制可能与促进P21cip1/WAF1表达、抑制Survivin的表达有关.     

Effect of carvedilol on Ca2+ movement and cytotoxicity in human MG63 osteosarcoma cells

Carvedilol is a useful cardiovascular drug for treating heart failure, however, the in vitro effect on many cell types is unclear. In human MG63 osteosarcoma cells, the effect of carvedilol on intracellular Ca2+ concentrations ([Ca2+]i) and cytotoxicity was explored by using fura-2 and tetrazolium, respectively. Carvedilol at concentrations greater than 1 microM caused a rapid rise in [Ca2+]i in a concentration-dependent manner (EC50=15 microM). Carvedilol-induced [Ca2+]i rise was reduced by 60% by removal of extracellular Ca2+. Carvedilol-induced Mn2+-associated quench of intracellular fura-2 fluorescence also suggests that carvedilol induced extracellular Ca2+ influx. In Ca2+-free medium, thapsigargin, an inhibitor of the endoplasmic reticulum Ca2+-ATPase, caused a monophasic [Ca2+]i rise, after which the increasing effect of carvedilol on [Ca2+]i was inhibited by 50%. Conversely, pretreatment with carvedilol to deplete intracellular Ca2+ stores totally prevented thapsigargin from releasing more Ca2+. U73122, an inhibitor of phospholipase C, abolished histamine (an inositol 1,4,5-trisphosphate-dependent Ca2+ mobilizer)-induced, but not carvedilol-induced, [Ca2+]i rise. Pretreatment with phorbol 12-myristate 13-acetate and forskolin to activate protein kinase C and adenylate cyclase, respectively, did not alter carvedilol-induced [Ca2+]i rise. Separately, overnight treatment with 0.1-30 microM carvedilol inhibited cell proliferation in a concentration-dependent manner. These findings suggest that in human MG63 osteosarcoma cells, carvedilol increases [Ca2+]i by stimulating extracellular Ca2+ influx and also by causing intracellular Ca2+ release from the endoplasmic reticulum and other stores via a phospholipase C-independent manner. Carvedilol may be cytotoxic to osteoblasts.     

Mechanisms of histamine-induced intracellular Ca2+ release and extracellular Ca2+ entry in MG63 human osteosarcoma cells

The effect of histamine on intracellular free Ca2+ levels ([Ca2+](i)) in MG63 human osteosarcoma cells was explored using fura-2 as a Ca2+ dye. Histamine increased ([Ca2+](i)) in a concentration-dependent fashion with an EC(50) value of 0.5 microM. Extracellular Ca2+ removal inhibited the ([Ca2+](i)) signals. Histamine failed to increase ([Ca2+](i)) in Ca2+-free medium after cells were pretreated with thapsigargin (an endoplasmic reticulum Ca2+ pump inhibitor). Addition of Ca2+ induced concentration-dependent ([Ca2+](i)) increases after preincubation with histamine in Ca2+-free medium. Histamine-induced intracellular Ca2+ release was abolished by inhibiting phospholipase C with 1-(6-((17beta-3-methoxyestra-1,3,5(10)-trien-17-yl)amino)hexyl)-1H-pyrrole-2,5-dione (U73122). The ([Ca2+](i)) increase induced by histamine in Ca2+ medium was abolished by cimetidine, but was not altered by pyrilamine, nifedipine, verapamil, and La(3+). Together, this study shows that histamine increased in ([Ca2+](i)) in osteosarcoma cells by stimulating H2 histamine receptors. The Ca2+ signal was caused by Ca2+ release from the endoplasmic reticulum in a phospholipase C-dependent manner. The Ca2+ release was accompanied by Ca(2+) influx.     

3,3'-Diindolylmethane alters Ca2+ homeostasis and viability in MG63 human osteosarcoma cells

Abstract: The effect of the natural product 3,3′‐diindolylmethane (DIM) on cytosolic Ca²⁺ concentrations ([Ca²⁺]_i) and viability in MG63 human osteosarcoma cells was explored. The Ca²⁺‐sensitive fluorescent dye fura‐2 was applied to measure [Ca²⁺]_i. DIM at concentrations of 40–80 μM induced a [Ca²⁺]_i rise in a concentration‐dependent manner. The response was reduced partly by removing Ca²⁺. DIM‐evoked Ca²⁺ entry was suppressed by nifedipine, econazole, SK&F96365 and protein kinase C modulators. In the absence of extracellular Ca²⁺, incubation with the endoplasmic reticulum Ca²⁺ pump inhibitors thapsigargin or 2,5‐di‐tert‐butylhydroquinone (BHQ) inhibited or abolished DIM‐induced [Ca²⁺]_i rise. Incubation with DIM also inhibited thapsigargin or BHQ‐induced [Ca²⁺]_i rise. Inhibition of phospholipase C with U73122 abolished DIM‐induced [Ca²⁺]_i rise. At concentrations of 10–50 μM, DIM killed cells in a concentration‐dependent manner. This cytotoxic effect was not altered by chelating cytosolic Ca²⁺ with 1,2‐bis(2‐aminophenoxy)ethane‐N,N,N′,N′‐tetraacetic acid (BAPTA). Annexin V/propidium iodide staining data implicate that DIM (20 and 40 μM) induced apoptosis in a concentration‐dependent manner. In sum, in MG63 cells, DIM induced a [Ca²⁺]_i rise by evoking phospholipase C‐dependent Ca²⁺ release from the endoplasmic reticulum and Ca²⁺ entry via protein kinase C‐sensitive store‐operated Ca²⁺ channels. DIM caused cell death that may involve apoptosis.     

Ca(2+) mobilization induced by W-7 in MG63 human osteosarcoma cells.

The effect of N-(6-aminohexyl)-5-chloro-1-naphthalenesulfonamide hydrochloride (W-7), a widely used calmodulin inhibitor, on intracellular free Ca(2+)levels ([Ca(2+)](i)) in MG63 human osteosarcoma cells was explored using fura-2 as a Ca(2+)probe. W-7 (20-1000 micro m) induced an increase in [Ca(2+)](i)in a dose-dependent manner, with an EC(50)of 100 microm. The [Ca(2+)](i)signal comprised an initial rise and a sustained plateau without significant decay within 5 min. External Ca(2+)removal decreased the Ca(2+)signals by reducing the peak and sustained phase, indicating W-7-activated intracellular Ca(2+)release and extracellular Ca(2+)influx. W-7 (500 microm) failed to induce a [Ca(2+)](i)increase in a Ca(2+)-free medium after pre-treatment with thapsigargin (1 microm), an endoplasmic reticulum Ca(2+)pump inhibitor. Conversely, W-7 pre-treatment abolished the Ca(2+)release induced by thapsigargin. This suggests that W-7 (500 microm ) released internal Ca(2+)mainly from the endoplasmic reticulum. The addition of 3 mm Ca(2+)increased [Ca(2+)](i)dose-dependently after preincubation with 20-1000 microm W-7 in a Ca(2+)-free medium, implying that W-7 induced capacitative Ca(2+)entry. W-7-induced Ca(2+)release was not altered by inhibiting phospholipase C with 2 microm 1-(6-((17 beta - 3-methoxyestra-1,3, 5(10)-trien-17-yl)amino)hexyl)-1H-pyrrole-2,5-dione) (U73122). Tryphan blue assay demonstrated that W-7 (200 microm) caused gradual cell death within 30 min of the initial drug exposure. Together, it was found that W-7 induced [Ca(2+)](i)increases in human osteosarcoma cells by releasing internal Ca(2+)from the endoplasmic reticulum, and also by triggering Ca(2+)influx. W-7 may be cytotoxic to osteosarcoma cells.     

紫金散粗提液抑制骨肉瘤细胞MG63恶性增殖的实验研究

目的探讨中药紫金散对人骨肉瘤细胞的杀伤作用。方法将紫金散粗提液与参鹿痨疽粗提液分别应用于体外培养的骨肉瘤MG63细胞,利用MTT法,流式细胞仪、倒置显微镜分别观察骨肉瘤细胞的生长抑制率、凋亡率以及细胞形态变化。观察其对肿瘤细胞的促凋亡作用。结果A组加入紫金散粗提液3d后骨肉瘤细胞凋亡率为6.57%,B组加入参鹿痨疽粗提液3d后骨肉瘤细胞凋亡率为18.24%,作用更明显。结论应用紫金散可明显促进肿瘤细胞的凋亡。     

Effect of protriptyline on [Ca2+]i and viability in MG63 human osteosarcoma cells

Tricyclic antidepressants (TCA) have been clinically prescribed in the auxiliary treatment of cancer patients. Although protriptyline, a type of TCA, was used primarily in the clinical treatment of mood disorders in cancer patients, the effect of protriptyline on physiology in human osteosarcoma is unknown. This study examined the effect of protriptyline on cytosolic free Ca^2+?concentrations ([Ca²⁺]_i) and viability in MG63 human osteosarcoma cells. Protriptyline between 50 and 250?μM evoked [Ca²⁺]_i rises concentration-dependently. Protriptyline induced influx of Mn²⁺, indirectly implicating Ca^2+?influx. Protriptyline-evoked Ca^2+?entry was inhibited by nifedipine by 20% but was not altered by econazole, SKF96365, GF109203X, and phorbol-12-myristate-13-acetate (PMA). In Ca²⁺-free medium, treatment with protriptyline inhibited the endoplasmic reticulum Ca^2+?pump inhibitor thapsigargin-evoked [Ca²⁺]_i rises. Conversely, treatment with thapsigargin inhibited 45% of protriptyline-evoked [Ca²⁺]_i rises. Inhibition of phospholipase C (PLC) with U73122 failed to alter protriptyline-evoked [Ca²⁺]_i rises. Protriptyline at 50–250?μM decreased cell viability, which was not reversed by pretreatment with the Ca^2+?chelator 1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid-acetoxymethyl ester (BAPTA/AM). Collectively, our data suggest that in MG63 cells, protriptyline induced [Ca²⁺]_i rises by evoking Ca^2+?release from the endoplasmic reticulum and other stores in a PLC-independent manner, and Ca^2+?entry via a nifedipine-sensitive Ca^2+?pathway. Protriptyline also caused Ca²⁺-independent cell death.     

The anti-anginal drug fendiline increases intracellular Ca(2+) levels in MG63 human osteosarcoma cells

The effect of fendiline, an anti-anginal drug, on cytosolic free Ca(2+) levels ([Ca(2+)](i)) in MG63 human osteosarcoma cells was explored by using fura-2 as a Ca(2+) indicator. Fendiline at concentrations between 1 and 200 microM increased [Ca(2+)](i) in a concentration-dependent manner and the signal saturated at 100 microM. The Ca(2+) signal was inhibited by 65+/-5% by Ca(2+) removal and by 38+/-5% by 10 microM nifedipine, but was unchanged by 10 microM La(3+) or verapamil. In Ca(2+)-free medium, pre-treatment with 1 microM thapsigargin (an endoplasmic reticulum Ca(2+) pump inhibitor) to deplete the endoplasmic reticulum Ca(2+) store inhibited fendiline-induced intracellular Ca(2+) release. The Ca(2+) release induced by 50 microM fendiline appeared to be independent of IP(3) because the [Ca(2+)](i) increase was unaltered by inhibiting phospholipase C with 2 microM U73122. Collectively, the results suggest that in MG63 cells fendiline caused an increase in [Ca(2+)](i) by inducing Ca(2+) influx and Ca(2+) release in an IP(3)-independent manner.     

Effect of acetaminophen (paracetamol) on human osteosarcoma cell line MG63

Aim:

To examine the effects of acetaminophen (paracetamol), a nonsteroidal anti-inflammatory drug (NSAID), on different cellular and functional parameters of the human osteosarcoma cell line MG63.

Methods:

Flow cytometry was used to study proliferation, antigenic profile, and phagocytic activity, and radioimmunoassay was used to determine osteocalcin synthesis as a cell differentiation marker.

Results:

Short-term treatment with therapeutic doses of paracetamol(5 or 25 μmol/L) reduced cell proliferation, osteocalcin synthesis, and phagocyte activity, and increased the expression of antigens involved in antigen presentation to T lymphocytes (CD80, CD86, HLA-DR).

Conclusion:

These findings suggest that paracetamol activates the osteoblast, inducing its immunogenic action to the detriment of its bone formation capacity.     

利多卡因调控微小RNA-195对骨肉瘤MG63细胞增殖、凋亡及化疗敏感性的影响

目的 探讨利多卡因(lidocaine)对骨肉瘤MG63细胞增殖、凋亡及化疗敏感性的影响及其可能的作用机制.方法 研究于2018年5―12月,体外培养骨肉瘤MG63细胞,不同浓度的利多卡因干预MG63细胞,采用甲基噻唑基四唑(MTT)法检测MG63细胞存活率及顺铂半抑制浓度(IC50值);流式细胞术检测细胞凋亡率;实时...     

姜黄素的抗骨肉瘤活性及其相关作用机理

目的探讨姜黄素对人成骨肉瘤MG-63细胞的抗肿瘤活性及其相关作用机制。方法 MTT实验检测姜黄素对人成骨肉瘤细胞MG-63的细胞毒作用;Annexin V/PI双染,流式细胞仪检测细胞凋亡的发生;提取细胞总蛋白Western blot实验检测细胞PARP蛋白的裂解带和survivin蛋白的表达变化。结果姜黄素对人成骨肉瘤MG-63细胞具有高效的细胞毒作用,其IC50值为(11.55±0.63)μmol/L;10,20和40μmol/L姜黄素处理MG-63细胞48 h后,细胞的凋亡率从(2.73±0.56)%依次升高到(18.1±1.22)%,(36.41±4.52)%和(57.23±5.86)%;Westernblot结果显示姜黄素诱发了MG-63细胞内PARP蛋白的裂解,同时姜黄素也可剂量依赖性的降低细胞内survivin蛋白的表达。结论姜黄素显示了高效的致骨肉瘤细胞凋亡活性,下调survivin蛋白可能是其诱导MG-63细胞凋亡的主要机制。     

Effects of Antrodia camphorata on viability,apoptosis, [Ca2+]i,and MAPKs phosphorylation in MG63 human osteosarcoma cells

The present study explored the effect of Antrodia camphorata (AC) on viability, apoptosis, mitogen‐activated protein kinases (MAPKs) phosphorylation, and Ca²⁺ regulation in MG63 human osteosarcoma cells. AC (25–50 µg/ml) did not affect cell viability, but at 100–200 µg/ml decreased viability and induced apoptosis in a concentration‐dependent manner. AC at concentrations of 25–200 µg/ml did not alter basal [Ca²⁺]_i, but at 25 µg/ml decreased [Ca²⁺]_i increases induced by ATP, bradykinin, histamine, and thapsigargin. ATP, bradykinin, and histamine increased cell viability while thapsigargin decreased it. AC (25 µg/ml) pretreatment failed to alter bradykinin‐ and thapsigargin‐induced effects on viability, but potentiated ATP‐ and histamine‐induced increases in viability. Immunoblotting showed that MG63 cells did not have background phospho‐JNK and phospho‐p38 mitogen‐activated protein kinases (MAPKs); and AC did not induce the phosphorylation of these two MAPKs. Conversely, the cells had significant background phospho‐ERK MAPK that was inhibited by 200 µg/ml AC. The ERK‐specific inhibitor PD98059 also induced cell death. Collectively, in MG63 cells, AC exerted multiple effects on viability and [Ca²⁺]_i, caused apoptosis probably via inhibition of ERK MAPK phosphorylation. Drug Dev Res 68:71–78, 2007. © 2007 Wiley‐Liss, Inc.     

Effects of MK‐886, a leukotriene synthesis inhibitor,on [Ca2+]i and apoptosis in MG63 human osteosarcoma cells

The effect of MK‐886 (3‐[1‐(p‐chlorobenzyl)‐5‐(isopropyl)‐3‐tert‐butylthioindol‐2‐yl]‐2, 2‐dimethylpropanoic acid), a compound widely used to inhibit leukotriene synthesis, on cytosolic free Ca²⁺ concentrations ([Ca²⁺]_i) in osteosarcoma cells has not been explored. This study examined whether MK‐886 altered [Ca²⁺]_i levels in suspended MG63 human osteosarcoma cells using fura‐2. MK‐886 at 0.1 μM and above increased [Ca²⁺]_i in a concentration‐dependent manner. The Ca²⁺ signal was reduced partly by removing extracellular Ca²⁺. MK‐886 induced Mn²⁺ quenching of fura‐2 fluorescence, implicating Ca²⁺ entry. MK‐886‐induced Ca²⁺ influx was inhibited by store‐operated Ca²⁺ entry inhibitors, nifedipine, econazole, and SKF96365; and by the protein kinase C modulators, phorbol 12‐myristate 13‐acetate (PMA) and GF109203X. In Ca²⁺‐free medium, after pretreatment with 5 μM MK‐886, 1 μM thapsigargin (an endoplasmic reticulum Ca²⁺ pump inhibitor)‐induced [Ca²⁺]_i rises were abolished; conversely, thapsigargin pretreatment nearly abolished MK‐886‐induced [Ca²⁺]_i rises. Inhibition of phospholipase C with U73122 did not change MK‐886‐induced [Ca²⁺]_i rises. Collectively, in MG63 osteosarcoma cells, MK‐886 induced [Ca²⁺]_i rises by causing phospholipase C‐independent Ca²⁺ release from the endoplasmic reticulum and Ca²⁺ influx via protein kinase C‐regulated store‐operated Ca²⁺ entry. Drug Dev Res 69: 49–57, 2008. © 2008 Wiley‐Liss, Inc.     

微小 RNA-490-3p靶向 FK506相关蛋白 14表达抑制骨肉瘤 MG63细胞侵袭和迁移的实验研究

目的探讨微小 RNA-490-3p(miR-490-3p)对骨肉瘤 MG63细胞侵袭、迁移的影响及其机制。方法 2019年 9月至 2020年 4月,从中科院上海细胞库购买人骨肉瘤细胞株 MG63进行体外培养,分为对照组(未转染)miR-NC组(转染 miR-NC)、 miR-490-3p组(转染 miR-490-3p mimics),miR-490-3p+pcDNA组(共转染 miR-490-3p mimics与空载体、)和 miR-490-3p+FKBP14组(共转染 miR-490-3p mimics与 FKBP14过表达载体),采用 RT-PCR检测 miR-490-3p表达, Transwell小室法检测细胞侵袭、迁移,蛋白质印迹法检测钙黏蛋白 E(E-cadherin)、基质金属蛋白酶 -2(MMP-2)、上皮间质转化因子 Twist转录因子( Twist)和 FK506相关蛋白 14(FKBP14)蛋白表达,双荧光素酶报告基因实验检测 miR-490-3p和 FKBP14的靶向关系。结果与对照组比较, miR-490-3p组细胞中 miR-490-3p表达水平明显升高( 3.68±0.37比 1.02±0.10,P<0.05),而侵袭细胞数( 33.25±2.02比     

肉桂酸对人骨肉瘤MG-63细胞增殖和分化的影响

目的研究中药有效成分肉桂酸对人骨肉瘤MG-63细胞增殖和分化的影响。方法肉桂酸处理MG-63细胞前后,台盼蓝拒染法进行活细胞计数;流式细胞仪检测细胞周期时相变化;光镜和电镜观察细胞形态和超微结构;VanGieson染色检测细胞Ⅰ型胶原表达;茜素红染色观察钙化骨结节形成;免疫细胞化学检测骨粘素和骨钙蛋白表达。结果肉桂酸明显抑制MG-63细胞增殖,D7抑制率达54.94%±4.69%。周期分析结果显示,肉桂酸处理后G0/G1期细胞比例较对照组明显升高(P<0.01)。光镜和电镜结果显示,肉桂酸处理组MG-63细胞形态和超微结构发生明显改变,呈现相应正常细胞的形态和超微结构特征。肉桂酸促进成骨细胞分化标志物Ⅰ型胶原、骨粘素和骨钙蛋白的表达与钙沉积以及典型骨结节的形成。结论肉桂酸可抑制人骨肉瘤MG-63细胞增殖并诱导其向成骨细胞分化。     

Polydatin induces apoptosis and autophagy via STAT3 signaling in human osteosarcoma MG-63 cells

Journal of Natural Medicines - Polydatin, a natural product, is detected in many daily diets, such as grape juices and peanut. Autophagy regulation is recognized as a new potential strategy for...     

Mechanisms of diethylstilbestrol-induced relaxation in rat aorta smooth muscle

The mechanisms of diethylstilbestrol (1 to 30 microM)-induced relaxation on noradrenaline (30 nM)-raised tone in the rat aorta smooth muscle were studied. Neither the increase of calcium content in the medium (3, 6 and 9 mM) nor Bay K 8644 (3, 10 and 100 nM) reversed diethylstilbestrol relaxation. Tamoxifen (3 microM), the quaternary derivate (tamoxifen ethyl bromide, 3 microM), actinomycin D (30 microM), cycloheximide (100 microM), Rp-cAMPS (30 microM), TPCK (1 microM) and difluoromethylornithine (1 mM) inhibited diethylstilbestrol-induced relaxation. Incubation with 2 microg/ml pertussis toxin, propranolol (1 microM), H-7 (10 microM), 2',3'- and 2',5'-dideoxiadenosine (10 and 30 microM, respectively) and methylene blue (10 microM) did not modify diethylstilbestrol-induced relaxation. Our results showed that presumably an activation of membrane mechanisms, protein kinase A activation, genomic mechanisms and polyamine synthesis might participate in diethylstilbestrol-elicited relaxation in addition to the increase in K(ATP) permeability, as previously described. Actinomycin D produces a synergistic effect, with tamoxifen, difluoromethylornithine and glibenclamide antagonizing the effect of diethylstilbestrol. In the case of the association of actinomycin D and glibenclamide, the antagonism of relaxation is complete. The fact that tamoxifen- and difluoromethylornithine-dependent mechanisms participate in diethylstilbestrol relaxation inhibited by glibenclamide suggests that two transduction pathways are involved in the relaxation. Therefore, K(ATP) channels and genomic mechanisms, both modulated by cyclic AMP (cAMP)-dependent mechanisms, are associated with diethylstilbestrol relaxation.     

Effect of calmidazolium on Ca(+2) movement and proliferation in human osteosarcoma cells

In human MG63 osteosarcoma cells, the effect of calmidazolium on [Ca(2+)](i) and proliferation was explored using fura-2 and ELISA, respectively. Calmidazolium, at concentrations greater than 0.1 micromol/L, caused a rapid increase in [Ca(2+)](i) in a concentration-dependent manner (EC(50) = 0.5 micromol/L). The calmidazolium-induced [Ca(2+)](i) increase was reduced by 66% by removal of extracellular Ca(2+). In Ca(2+)-free medium, thapsigargin, an inhibitor of the endoplasmic reticulum Ca(2+)-ATPase, caused a monophasic increase in [Ca(2+)](i), after which the effect of calmidazolium to increase [Ca(2+)](i) was completely inhibited. U73122, an inhibitor of phospholipase C (PLC), abolished histamine (but not calmidazolium)-induced increases in [Ca(2+)](i). Pretreatment with phorbol 12-myristate 13-acetate to activate protein kinase C inhibited the calmidazolium-induced increase in [Ca(2+)](i) in Ca(2+)-containing medium by 47%. Separately, it was found that overnight treatment with 2-10 micromol/L calmidazolium inhibited cell proliferation in a concentration-dependent manner. These results suggest that calmidazolium increases [Ca(2+)](i) by stimulating extracellular Ca(2+) influx and also by causing release of intracellular Ca(2+) from the endoplasmic reticulum in a PLC-independent manner. Calmidazolium may be cytotoxic to osteosarcoma cells.     

四丁基丙二胺对人MG63骨髓瘤细胞增殖、凋亡和迁移能力的影响

目的分析新多胺类似物四丁基丙二胺(tetrabutylpropanediamine,TBP)对人骨髓瘤MG63细胞生长的影响及其机制。方法 MTT法用于分析细胞增殖,流式细胞分析用于细胞周期和凋亡细胞鉴定,琼脂糖凝胶电泳用于分析DNA片段化,Western blot用于分析细胞凋亡相关蛋白的表达水平,Transwell技术用于分析细胞的迁移能力。结果TBP明显抑制MG63细胞的增殖和迁移能力,抑制效应呈时间和剂量依赖性。流式细胞分析发现,TBP干扰细胞周期,导致G1和G2期细胞比例升高,但S期细胞比例明显下降,同时凋亡细胞数量大幅增加。TBP处理后,细胞染色体DNA出现凋亡细胞典型的DNA片段化现象,细胞质中促凋亡蛋白Bax和细胞色素C含量明显升高。结论 TBP具有抑制人骨髓瘤MG63细胞增殖的药理活性,其机制与抑制细胞周期和诱导细胞凋亡有关,提示TBP具有用于临床骨髓瘤治疗的潜在价值。