卵巢浆液性囊腺癌组织中 miR-20a、Let-7a表达变化及与患者预后的关系

目的观察卵巢浆液性囊腺癌组织中微小RNA-20a(miR-20a)和Let-7a的表达变化,并探讨二者与患者预后的关系。方法卵巢浆液性囊腺癌患者93例,术中留取肿瘤组织。分别留取同期手术治疗的卵巢交界性肿瘤患者(43例)及卵巢浆液性囊腺瘤患者(48例)的肿瘤组织,留取正常卵巢组织(40例份)。采用real-time PCR技术检测不同组织中的miR-20a和Let-7a。采用Kaplan-Meier法分析卵巢浆液性囊腺癌组织中miR-20a、Let-7a表达与患者预后的关系。结果各种卵巢肿瘤组织中miR-20a表达量高于正常卵巢组织、Let-7a表达量低于正常卵巢组织(P均<0.05);卵巢浆液性囊腺癌组织中miR-20a表达量高于卵巢交界性肿瘤组织及卵巢浆液性囊腺瘤组织、Let-7a表达量低于卵巢交界性肿瘤组织及卵巢浆液性囊腺瘤组织(P均<0.05)。miR-20a、Let-7a与卵巢浆液性囊腺癌的临床分期、分化程度、淋巴结转移和远处转移发生情况有关(P均<0.05)。卵巢浆液性囊腺癌组织中miR-20a与Let-7a表达呈负相关关系(r=-0.489,P<0.05)。卵巢浆液性囊腺癌患者随访至2014年2月28日,miR-20a表达量>1.42者中位生存时间短于miR-20a表达量≤1.42者,Let-7a表达量<0.98者中位生存时间短于Let-7a表达量≥0.98者(P均<0.05)。结论卵巢浆液性囊腺癌组织中miR-20a呈高表达、Let-7a呈低表达,二者可能共同参与了卵巢浆液性囊腺癌的发生、发展,并与患者预后有关。     

Let-7d*在增生性血管疾病中的表达及其作用机制

目的 探究Let-7d*在增生性血管疾病中的表达及其相关作用机制.方法 分别构建球囊损伤模型与平滑肌细胞增殖模型,研究Let-7d*在这两种模型中的表达趋势,并调控Let-7d*的表达,研究其相关生物学功能.结果 在球囊损伤诱导新生内膜生长的模型中,Let-7d*的表达水平较在正常颈动脉中明显降低,同时在增殖的血管平滑...     

Bmi-1-siRNA对乳腺癌MCF-7细胞体外增殖的抑制作用

目的探讨Bmi-1-siRNA对乳腺癌MCF-7细胞体外增殖的作用及机制。方法将不同浓度的Bmi-1-siRNA转染MCF-7细胞,MTT法检测细胞增殖情况,免疫细胞化学技术检测转染72h后MCF-7细胞内Bmi-1、端粒酶逆转录酶（hTERT）、细胞增殖相关核抗原Ki-67蛋白的表达。结果Bmi-1-siRNA对MCF-7细胞的生长抑制率明显升高,且随浓度增高而升高（P〈0．05）;MCF-7细胞中Bmi-1、hTERT、Ki-67蛋白表达明显降低（P〈0．05）。结论Bmi-1-siRNA可有效抑制MCF-7细胞的增殖,其机制为降低Bmi-1 mRNA及Bmi-1、hTERT蛋白的表达。     

藤黄酸对乳腺癌细胞MCF-7细胞端粒酶活性及其逆转录酶hTERT mRNA表达的抑制作用研究

目的研究藤黄酸对人乳腺癌细胞MCF-7端粒酶活性和端粒酶逆转录酶(hTERT)mRNA表达及其转录调控基因C-myc的影响,以探讨藤黄酸对人乳腺癌细胞端粒酶活性的调控机制。方法藤黄酸作用于人乳腺癌细胞24h、48h、72h后,收集细胞,四甲基偶氮唑盐(MTT)比色法检测MCF-7细胞生长抑制率,同时采用PCR-ELISA法检测端粒酶活性、RT-PCR法检测hTERT mRNA的表达、Western blot技术检测C-myc蛋白的表达。结果藤黄酸能明显抑制人乳腺癌细胞MCF-7生长和增埴,对端粒酶活性、hTERT mRNA及转录调控基因C-myc的表达具有抑制作用,并呈明显的时效和量效相关性。结论藤黄酸能够明显抑制人乳腺癌细胞MCF-7增殖,其机制可能是对MCF-7细胞的端粒酶活性及其端粒酶转录调控基因C-myc有较好的抑制作用,提示藤黄酸有可能成为高效低毒的抗肿瘤天然药物。     

钯配合物对人乳腺癌细胞MCF-7增殖的抑制作用及机制

目的 探讨钯配合物(PBIPS)对人乳腺癌细胞(MCF-7)增殖的抑制作用及其机制.方法 MTT法检测PBIPS对MCF-7的细胞毒性,荧光显微镜观察细胞形态学变化,流式细胞术(FCM)检测MCF-7细胞的凋亡率;Western-blotting检测不同浓度PBIPS作用48 h凋亡蛋白survivin 、caspase-3及bcl-2和bax的表达情况.结果 PBIPS可以时间和剂量依赖性地抑制MCF-7细胞的生长;与对照组比较,PBIPS不同浓度剂量组AO/EB荧光染色出现典型的凋亡形态学改变;FCM检测表明MCF-7细胞凋亡率各实验组均明显高于对照组(P＜0.01),且呈时间剂量依赖性;Western blotting显示与对照组相比,各实验组PBIPS可显著下调MCF-7细胞survivin和bcl-2的表达(P＜0.01),而caspase-3和bax的表达显著增加(P＜0.01).结论 PBIPS能明显诱导人乳腺癌MCF-7细胞凋亡,其机制可能与改变凋亡相关基因survivin、bcl-2、caspase-3及bax的表达有关.     

Ad—PTEN对乳腺癌细胞系MCF-7生长抑制作用的影响及机制

目的探讨携带野生型PTEN基因的重组腺病毒（Ad—PTEN）和LY294002对乳腺癌细胞系MCF-7生长抑制作用的影响及机制。方法在MCF-7中导人Ad—PTEN和P13K抑制剂LY294002。Westem blotting法检测P11EN／P13K／AKT及其下游相关蛋白表达,M1Tr法检测MCF-7细胞存活率,流式细胞仪检测细胞周期,AnnexinV法检测细胞凋亡率。结果与DMSO组、空载病毒和LY294002组相比,联合组（LY294002＋Ad—PTEN）PTEN蛋白明显上调,而P13K／AKT及下游蛋白水平显著下降;联合组细胞阻滞于G0／C1期;细胞凋亡率增加明显;自培养第2天起,联合组细胞生存率呈下降趋势。结论Ad—PTEN联合LY294002通过阻滞细胞周期、促进细胞凋亡抑制MCF-7的增殖能力,两者具有正向协同作用;其机制可能与下调P13K／AKT信号通路下游蛋白有关。     

上调乳腺癌细胞MCF-7中IGFBP-6基因对细胞增殖的影响

目的 探究IGFBP-6对人乳腺癌MCF-7增殖的影响.方法 收集经手术切除并经病理证实的乳腺癌组织标本80例及癌旁组织50例;体外培养乳腺癌细胞MCF-7;免疫组织化学法检测乳腺癌组织中IGFBP-6表达,将IGFBP-6质粒转染到MCF-7细胞中,MTT法观察上调IGFBP-6对MCF-7细胞增殖的影响,实时荧光定...     

MicroRNA-16及Let-7a在支气管哮喘患者中表达水平及与严重程度的关系

目的探讨微RNA-16(microRNA-16,miR-16)及Let-7a在支气管哮喘患者中表达水平及与严重程度的关系。方法选取2017年7月至2019年7月我院收治的128例支气管哮喘患者作为研究对象,按照严重程度将患者分为轻、中度组(n=85)和重度组(n=43)。选取同期120例体检健康者作为对照组。采用实时荧光定量PCR(RT-qPCR)检测血清miR-16和Let-7a相对表达量,并分析模型A(miR-16+Let-7a)、模型B(IL-13+IgE+EOS计数)及模型C(miR-16+Let-7a+IL-13+IgE+EOS计数)评估重度支气管哮喘的效能。结果轻、中度组及重度组血清miR-16和Let-7a相对表达量低于对照组,重度组血清miR-16和Let-7a相对表达量低于轻、中度组,差异均有统计学意义(P0.05)。miR-16、Let-7a评估重度支气管哮喘的AUC分别为0.802、0.808。Logistic回归分析显示IL-13、IgE、EOS计数、miR-16、Let-7a与支气管哮喘病情加重密切相关。模型C评估重度支气管哮喘的效能高于模型A及模型B。结论血清miR-16和Let-7a与支气管哮喘严重程度关系密切,检测血清miR-16和Let-7a相对表达量有助于评估患者病情。     

绿原酸对MCF-7细胞增殖的影响及机制探讨

目的为绿原酸（CGA）用于乳腺癌的治疗提供依据。方法将不同浓度的CGA干预乳腺癌MCF-7细胞;采用四甲基偶氮唑蓝（MTT）法检测CGA对MCF-7细胞的生长抑制率,流式细胞术检测MCF-7细胞凋亡率、细胞周期及细胞周期素（Cydin）D1表达。结果0．25、0．5mg／mlCGA作用MCF-7细胞48h后,使细胞阻滞于G1／G0期,两组的G1／G0期细胞较对照组明显增加（P〈0．01）;0、0．25、0．5mg／mlCGA处理MCF-7细胞48h后,Cyc-linD1的平均荧光强度比（MFIR）分别为9．64±0．18、9．15±0．22、8．104-0．28（P=0．001）。结论CGA可抑制MCF-7细胞增殖,使细胞阻滞于G0／G1期;其机制可能与下调CyclinD1表达有关。     

转染Zbtb7a基因对人胃癌SGC7901细胞增殖、凋亡的影响及其机制

目的 观察转染Zbtb7a基因对人胃癌细胞系SGC7901增殖及凋亡的影响,并探讨其机制.方法 将pcDNA3.1-Zbtb7a和pSilencer 3.1-H1-mk用Lipofectamine2000转染SGC7901细胞,采用RT-PCR和Western Blot法检测MK mRNA及蛋白表达,CCK-8试剂盒和平板克隆法检测细胞增殖,流式细胞仪Annexin V-PI染色检测细胞凋亡.结果 转染Zbtb7a后,SGC7901细胞中MK表达水平明显升高,细胞增殖能力明显增强（P＜0.05）,并且0.5ng/mL TRAIL诱导的细胞凋亡受到抑制（P＜0.05）.Zbtb7a高表达后干扰MK的表达,细胞增殖及克隆能力则明显降低（P＜0.05）,TRAIL诱导的细胞凋亡数也明显增加（P＜0.05）.结论 Zbtb7a可以通过上调MK的表达,促进SGC-7901细胞增殖以及抑制TRAI诱导的细胞凋亡.     

Effects of antioestrogens on the proliferation of MCF-7 human breast cancer cells

Non-steroidal antioestrogens, such as tamoxifen, inhibit the growth of human breast cancer cells. The experiments described here compare and contrast the efficacy of tamoxifen and the 'pure' antioestrogen, ICI 164384, on the inhibition of proliferation of MCF-7 cells. Previous studies have shown that ICI 164384 has a greater maximal inhibitory effect than conventional antioestrogens on the growth of MCF-7 cells. Both types of compound block progression of cells through the cell cycle in the early G1 phase. These studies have been extended to measure the population distribution of antioestrogen-treated cells by the use of two-parameter flow cytometry. ICI 164384 proved to be more effective than tamoxifen in decreasing the proportion of actively growing cells in an asynchronous population. In cells grown in the complete absence of exogenous oestrogens, growth was stimulated by oestradiol, insulin, insulin-like growth factor-I (IGF-I) or transforming growth factor-alpha (TGF-alpha). The potent metabolite of tamoxifen, trans 4'-hydroxytamoxifen (4'-OHT), alone also stimulated growth, whereas ICI 164384 did not. Oestradiol and insulin added together demonstrated a clear synergistic enhancement of cell growth. Correspondingly, the stimulatory effect of 4'-OHT on growth was magnified in the presence of insulin, and a combination of ICI 164384 with insulin revealed a much weaker stimulatory action of the 'pure' antagonist. For both compounds the interaction with insulin was complex and characterized by a bell-shaped dose-response curve. However, for 4'-OHT at all concentrations in the range 1 pM-1 microM in the presence of insulin, cell numbers were greater than in cultures exposed to insulin alone. This was not the case for ICI 164384 which suggested that differences in efficacy may be due to interactions between oestrogen and growth factor-mediated mechanisms. Furthermore, ICI 164384 was more effective in inhibiting the action of IGF-I and TGF-alpha alone or in combination, although both antioestrogens produced a partial blockade of growth factor responses in the complete absence of oestradiol. It is concluded that the difference in efficacy between partial agonist and 'pure' antagonist antioestrogens to inhibit growth in vitro is consistent with the difference in the pharmacological profile of these compounds. The absence of stimulatory activity of ICI 164384 is of particular significance in reducing to a minimum the synergistic interaction between oestrogens and insulin.(ABSTRACT TRUNCATED AT 400 WORDS)     

Melatonin effects on intercellular junctional communication in MCF-7 human breast cancer cells

Melatonin exerts a direct antiproliferative effect on estrogen-responsive MCF-7 cells in culture. Recently, the importance of the anti-invasive actions of melatonin as a part of the oncostatic action of this indolamine has been reported. Gap junctional intercellular communication is known to be involved in controlling cell proliferation and differentiation, and a decrease in intercellular junctional communication has been described in highly invasive mammary cancer cells. Because melatonin at physiological doses (1 nM) shifts MCF-7 cells to a lower invasive status, we postulate that melatonin could modulate the levels of gap junctional intercellular communication in these tumor cells. To test our hypothesis, we studied gap junctional intercellular communication in MCF-7 human breast cancer cells previously (7-8 days) treated, or not, with melatonin (10 microM or 1 nM). Using the scrape-loading assay dye-transfer technique to introduce 0.05% Lucifer yellow into cells, we measured the ability of the tumor cells to transfer dye to adjacent cells. Rhodamine dextran (0.05%) was used as a control dye to verify that dye-transfer occurs through intercellular junctions. The presence of melatonin (10 microM or 1 nM) in the culture medium significantly increased (P < 0.01) the transfer of the dye to adjacent cells through gap junctions. This increase was greater at 10 microM melatonin, and averaged scan profiles of cells treated with melatonin 10 microM showed a statistically significant increase (P < 0.01) in the integrated optical density values, and a broadening of the densitometric scan. These findings suggest that melatonin could exert its antitumor action, at least in part, by increasing regulatory signals that are passed between adjacent epithelial cells through intercellular junctions.     

89Sr照射对乳腺癌MCF-7细胞分泌sFas、sFasL的影响

目的 探讨89Sr照射对乳腺癌MCF-7细胞分泌sFas、sFasL的影响.方法 采用不同浓度89Sr照射乳腺癌MCF-7细胞,培养24h后,用酶联免疫吸附试验检测细胞培养上清液中的sFas、sFasL水平.结果 乳腺癌MCF-7细胞分泌sFas、sFasL;89Sr照射可明显抑制MCF-7细胞增殖,随着89 Sr浓度升高,其sFas、sFasL水平逐渐下降,89Sr不同浓度间的sFas、sFasL水平两两比较,P均＜0.05.结论 89Sr可诱导乳腺癌MCF-F细胞凋亡,89Sr照射能有效降低MCF-7细胞分泌sFas、sFasL的水平;sFas的动态变化可能与乳腺癌细胞发生、发展的病理生理过程有关.     

Modulation of intracellular calcium and calmodulin by melatonin in MCF-7 human breast cancer cells

The pineal hormone, melatonin, has been shown to inhibit the proliferation of the estrogen receptor alpha (ERalpha)-positive macrophage chemotactic factor (MCF)-7 human breast cancer cells. Previous studies from other systems indicate that melatonin modulates the calcium (Ca2+)/calmodulin (CaM) signaling pathway either by changing intracellular calcium concentration ([Ca2+]i) via activation of its G-protein coupled membrane receptors, or through a direct interaction with CaM. In this study, although melatonin alone had no effect on basal [Ca2+]i in MCF-7 cells, it significantly enhanced the elevation of [Ca2+]i induction by extracellular adenosine triphosphate (ATP), which increases [Ca2+]i via the G protein-coupled P2y-purinoceptor and the phospholipase C (PLC) pathway. Pretreatment of MCF-7 cells with 10(-7) M melatonin increased the 10(-5) M ATP-induced [Ca2+]i peak change from 79.4 +/- 11.6 nM to 146.2 +/- 22.3 nM. Furthermore, without changing total cellular CaM levels, melatonin markedly increased the amount of membrane-bound CaM to 237 and 162% of control levels after I and 6 hr of treatment, respectively. Cytosolic CaM levels were also elevated to 172% of control after 6 hr of melatonin treatment. Correlative growth studies demonstrated that ATP (10(-5) M) can stimulate MCF-7 cell growth, that melatonin can suppress MCF-7 cell proliferation, but that pretreatment of MCF-7 cells with melatonin followed by ATP(10(-5) M), like 10(-4) M ATP can further suppress MCF-7 cell proliferation; this indicates that melatonin's potentiation of ATP induced [Ca2+]i may be above the threshold for cell growth. Given the important role of [Ca2+]i and CaM in tumor cell homeostasis and proliferation and melatonin's modulation of [Ca2+]i, melatonin's effects on the Ca2+/CaM signaling pathway may play an important role in mediating the growth-inhibitory effect of melatonin on MCF-7 human breast cancer cells.