erbB-3结合蛋白ebp1对Tca8113细胞生长增殖的影响

目的探讨erbB3结合蛋白ebp1对口腔鳞状细胞癌Tca8113细胞生长增殖的影响。方法免疫组化筛选erbB2/erbB3双阳性的口腔鳞状细胞癌Tca8113细胞系;转染pcDNA3.1ebp1质粒至Tca8113细胞,G418筛选阳性细胞;RTPCR检测转染后细胞中ebp1的转录水平;通过细胞生长曲线、3HTdR摄入量以及集落形成率观察转染前后细胞生长增殖能力的改变。结果Tca8113细胞同时表达erbB2/erbB3;RTPCR显示转染后细胞中ebp1转录水平明显升高;Tca8113ebp1细胞生长曲线平缓,生长减慢;3HTdR摄入量明显减少,细胞增殖能力减弱。结论ebp1在体外能显著抑制Tca8113肿瘤细胞的生长增殖。

胃腺癌组织中circ-104533的表达情况及其对胃癌细胞生长增殖的影响

目的探讨胃腺癌组织中环状RNA-104533(circ-104533)的表达情况及对其对胃癌细胞生长增殖的影响。方法通过circRNA芯片得出在胃癌中有统计学差异的11个高表达circRNA,然后经过qRT-PCR实验在78例胃腺癌组织中进行验证,仅得出circ-104533和circ-102712呈现高表达水平。本研究以circ-104533为研究对象,分析胃腺癌组织中circ-104533表达水平与临床病理特征的相关性。通过体外细胞转染技术建立过表达circ-104533和敲低表达circ-104533表达的胃癌SGC-7901细胞株,采用CCK-8法和平板克隆实验检测胃癌细胞的生长和增殖能力。结果胃腺癌组织中circ-104533的表达水平高于癌旁正常组织(P<0.05)。根据circ-104533表达水平将胃腺癌组织标本分为正常/低表达组(19例)和高表达组(59例)。circ-104533表达水平与肿瘤大小和分化程度密切相关(P<0.05),而与患者年龄、性别、肠化、病理类型、淋巴结转移和TNM分期无相关性(P>0.05)。体外细胞实验表明,过表达circ-104533后癌细胞72 h OD值和克隆形成数大于正常癌细胞组(P<0.05),而敲低表达circ-104533后癌细胞72 h OD值和克隆形成数小于正常癌细胞组,差异具有统计学意义(P<0.05)。结论胃腺癌组织中circ-104533表达水平升高,其可能是胃腺癌一种新的促癌基因;circ-104533能促进胃腺癌的发生发展过程。

Irisin促进上皮性卵巢癌细胞生长增殖

目的Irisin是近几年新发现的一种与能量代谢、胰岛素抵抗等相关的细胞因子,在人体中广泛表达且与多种肿瘤的病理过程密切相关,本研究旨在明确Irisin对上皮性卵巢癌细胞A2780是否有促增殖作用。方法用不同浓度的Irisin处理细胞A2780,通过CCK-8实验检测细胞生长增殖情况。结果CCK-8实验表明Irisin能促进细胞生长增殖。结论Irisin能诱导卵巢癌细胞生长增殖,进一步促进肿瘤发生发展。

凝血酶敏感蛋白-1对胆囊癌细胞生长增殖的作用

目的探讨研究凝血酶敏感蛋白-1(TSP-1)对胆囊癌细胞株GBC-SD生长增殖的作用及其作用途径。方法根据处理因素将胆囊癌细胞分为:空白对照组,TSP-1组,CD36单抗阻断组、CD47单抗阻断组。用四唑盐比色法(MTT)和流式细胞术检测TSP-1及受体CD36、CD47对胆囊癌细胞生长、细胞周期的影响。结果TSP-1组、CD36阻断组、CD47阻断组细胞生长明显低于对照组(抑制率分别为:23.63%,9.57%,18.13%)。CD36阻断组的抑制率小于CD47阻断组。TSP-1组G0/G1期细胞数增加,S期和G2/M期细胞数减少,与对照组和CD36阻断组相比有显著性差异。增殖指数分别为:对照组(39.55±2.12)、TSP-1组(26.21±0.61)、CD36阻断组(36.43±1.28)、CD47阻断组(30.73±0.38);各组之间两两比较均有差异。结论TSP-1可对抑制胆囊癌细胞的生长,通过与受体CD36的结合阻滞胆囊癌细胞停止于G0/G1期可能是主要的作用途径。

脾虚痰浊巴马小型猪冠脉细胞凋亡与细胞生长和增殖相关基因差异性表达研究

目的:基于PCR array技术观察脾虚痰浊巴马小型猪冠脉组织细胞凋亡和生长与增殖相关基因差异性表达,探讨AS发生的可能机制。方法:在建立脾虚痰浊巴马小型猪模型基础上,应用PCR array技术检测巴马小型猪冠脉细胞增殖与凋亡的相关基因的表达差异性表达。结果:PCR array显示巴马小型猪冠脉细胞中与细胞凋亡相关基因BAX、BCL2、TGFβ1、TNF、BCL2L1及与细胞生长和增殖相关的基因CSF-2、CCL2、MMP1、MMP3、FGF2均发生差异性变化。结论:脾虚痰浊巴马小型猪冠脉细胞增殖与凋亡相关基因的变化可能是脾虚痰浊冠状动脉硬化发生的病理机制。

FSH对体外培养猪卵巢颗粒细胞生长及增殖的影响

采集成年健康母猪的卵巢,收集卵泡颗粒细胞,在配制好的培养液中培养。颗粒细胞原代培养体系建立成功后,分别将不同质量浓度的FSH(0.1,0.5,1,5,25 ng/mL)作用于细胞,每隔48 h换1次培养液,加入新的FSH,倒置显微镜下检测细胞的生长及增殖状况。结果表明,FSH质量浓度越大,细胞生长状况越好,差异有统计学意义(P<0.05);在固定质量浓度的FSH作用下,细胞每次换液后生长状况都会变好。FSH能促进猪卵泡颗粒细胞增殖,并与作用时间长短有关。

青蒿油乳抗人肝癌细胞增殖生长抑制实验

目的 :探讨青蒿油乳抗肿瘤作用。方法 :采用MTT法检测青蒿油乳对人肝癌SMMC - 772 1细胞株的杀伤作用。结果 :三次重复实验表明 ,青蒿油乳浓度在 2 5 0 μg/ml时 ,对肝癌细胞最高抑制率达 87.0 % ,其IC50 为175 μg/ml。结论 :青蒿油乳有抗肝癌细胞的作用。

联合应用c-Ha-ras,c-myc反义寡聚脱氧核苷酸对人膀胱癌细胞生长增殖的抑制作用

据《中华外科杂志》1996年1月34卷 第1期报道 中国人民解放军兰州军区第三医院泌尿外科李鸣等,联合应用人工合成的c-H-ras,c-myc反义寡聚脱氧核苷酸(ASO)。

《肝细胞的生长、增殖和凋亡》

细胞渭亡(apoptosis)是一种生理现象，但也可见于病理过程，因此广泛地发生于各个生理的或是病理的器官组织。为了更好地了解肝脏疾病的病理发生，2000年底，在日本的宇部召开了山口肝脏病学讨论会第12届年会，其主题是肝细胞的分化、坦殖与渭亡的信号系统调控。与会者皆为肝脏病学的世界著名科学家。本书即这次年会的精彩论文汇编。其主要论题有：一个与肝脏特异基因表达和肝脏干细胞相关的显性抑制性螺旋—环—螺旋蛋白—HHM；肝炎病人肝组织中的一种神经干细胞RNA结合蛋白—抗原musashi—1的表达；利用肝脏干细胞重组肝组织以及NF—κB对TNF—α和Fas诱导的肝细胞凋亡的调控；维甲酿诱导的肝细胞癌的凋亡—“克隆缺失”治疗(Clonal Deletion Therapy)的分子基础；正常肝细胞和肝癌细胞的细胞周期信号传导；多种生长因子同时应用对大昆肝细胞DNA合成、MAPK活性和G1周期蛋白的作用；p21WAFl／CIPl参与人肝癌细胞系Troglitazone诱导的细胞周期停滞；配体在肝细胞癌免疫发病机制中的作用；丙型肝炎病毒增殖相。

USP22截短质粒构建鉴定及其在肝细胞肝癌细胞中的功能初探

目的:构建编码ubiquitin specific peptidase 22(USP22)截短蛋白的表达质粒,探索USP22在肝细胞肝癌(hepatocellular carcinoma,HCC)中的作用及其分子机制,并初步解析USP22在HCC细胞系中对细胞增殖及索拉菲尼作用的影响。方法:设计合成针对于人源USP22截短质粒序列的引物,通过PCR的方法以含有USP22全长蛋白编码序列的表达载体作为模板,扩增目的片段,再用限制性内切酶EcoRⅠ和XhoⅠ进行双酶切,连接转化后,将构建好的质粒进行酶切鉴定及测序,证明USP22截短质粒构建成功。将构建好的USP22全长及截短表达质粒转染到HCC细胞系HCC-LM3中,分别进行Western blot和confocal实验检测USP22全长及截短质粒在HCC-LM3细胞中的表达及定位。生物学功能方面,利用MTS法检测USP22全长对HCC细胞增殖及索拉菲尼作用的影响。结果:成功构建了USP22截短表达质粒,并在HCC细胞系HCC-LM3中验证了其蛋白表达;USP22全长及所构建的截短质粒编码蛋白在HCC-LM3中定位于细胞核;此外,生物学功能实验证实USP22促进HCC-LM3细胞增殖,并削弱了索拉菲尼对HCC-LM3细胞生长的抑制作用。结论:在HCC-LM3细胞中,USP22及其截短质粒编码蛋白均定位于细胞核;USP22促进HCC细胞增殖,且可能参与在HCC治疗中的索拉菲尼耐受进程。

人类脂肪来源成体干细胞体外培养的生物学特征研究及供体年龄对其增殖的影响

通过研究人类脂肪组织来源成体干细胞(human adipose tissue derived adult stem cells,hADAS细胞)体外培养的生物学特征及供体年龄对其生长增殖的影响,为其作为组织工程学研究和应用的种子细胞提供实验室研究基础.取不同年龄组志愿者(<20岁、21～40岁、41～60岁和>61岁),手术中切除少量皮下脂肪组织,酶消化法分离细胞,体外培养,传至20代.取培养第3代细胞,流式细胞仪检测细胞表面抗原标记物(CD29,CD34,CD44,CD45,CD49d,HLA-DR,CD106)和细胞周期.细胞培养传20代后进行细胞染色体组分析.MTT法分别检测各年龄组不同时间点吸光值,绘制生长曲线,根据公式TD=t(lg2/lgNt-lgN_0)换算人类脂肪来源干细胞倍增时间(timedoubling,TD).结果显示:酶消化法自皮下脂肪组织分离细胞,体外培养hADAS细胞,流式细胞仪检测示CD29+,CD34-,CD44+,CD45-,CD49d±,HLA-DR-,CD106-;G_1期占90%(P10代);P20细胞染色体组分析显示为正常染色体,未见异常染色体,保持遗传稳定.绘制细胞生长曲线,<20年龄组TD为(11.22±0.73)h,21～40年龄组TD为(11.14±0.76)h,两组之间P>0.05,无显著性差异;41～60年龄组TD为(13.18±1.58)h,>61年龄组TD为(13.11±0.83)h,两组之间P>0.05,无显著性差异.<20年龄组TD与>61年龄组TD显著性差异,P<0.05.说明hADAS细胞可方便地取材于皮下脂肪组织,体外培养遗传特性稳定,生长活跃,青年组生长活性强于老年组,可以作为组织工程学种子细胞的新来源.

Long-term omeprazole and esomeprazole treatment does not significantly increase gastric epithelial cell proliferation and epithelial growth factor receptor expression and has no effect on apoptosis and p53 expression

AIM： To study the effect of proton pump inhibitor （PPI） treatment on patients with reflux esophagitis and its in vivo effect on apoptosis, p53- and epidermal growth factor receptor （EGFR） expression. METHODS： After informed consent was obtained, gastric biopsies of the antrum were taken from patients with reflux oesophagitis prior to and after 6 mo of 20 mg omeprazole （n = 24） or 40 mg esomeprazole （n = 22） therapy. Patients did not take any other medications known to affect the gastric mucosa. All patients were Helicobacter pylori negative as confirmed by rapid urease test and histology, respectively. Cell proliferation, apoptosis, EGFR, and p53 expression were measured by immunohistochemical techniques. At least 600 glandular epithelial cells were encountered and results were expressed as percentage of total cells counted. Was considered statistically significant. RESULTS： Although there was a trend towards increase of cell proliferation and EGFR expression both in omeprazole and esomeprazole treated group, the difference was not statistically significant. Neither apoptosis nor p53 expression was affected. CONCLUSION： Long-term PPI treatment does not significantly increase gastric epithelial cell proliferation and EGFR expression and has no effect on apoptosis and p53 expression.

Baicalin inhibits PDGF-BB-stimulated vascular smooth muscle cell proliferation through suppressing PDGFRβ-ERK signaling and increase in p27 accumulation and prevents injury-induced neointimal hyperplasia

The increased proliferation and migration of vascular smooth muscle cells （VSMCs） are key events in the development of atherosclerotic lesions. Baicalin, an herb-derived flavonoid compound, has been previously shown to induce apoptosis and growth inhibition in cancer cells through multiple pathways. However, the potential role of baicalin in regulation of VSMC proliferation and prevention of cardiovascular diseases remains unexplored. In this study, we show that pretreatment with baicalin has a dose-dependent inhibitory effect on PDGF-BB-stimulated VSMC pro- liferation, accompanied with the reduction of proliferating cell nuclear antigen （PCNA） expression. We also show that baicalin-induced growth inhibition is associated with a decrease in cyclin E-CDK2 activation and increase in p27 level in PDGF-stimulated VSMCs, which appears to be at least partly mediated by blockade of PDGF recep- tor [~ （PDGFR~）-extracellular signal-regulated kinase 1/2 （ERK1/2） signaling. In addition, baicalin was also found to inhibit adhesion molecule expression and cell migration induced by PDGF-BB in VSMCs. Furthermore, using an animal carotid arterial balloon-injury model, we found that baicalin significantly inhibited neointimal hyperplasia. Taken together, our results reveal a novel function of baicalin in inducing growth arrest of PDGF-stimulated VSMCs and suppressing neointimal hyperplasia after balloon injury, and suggest that the underlying mechanism involves the inhibition of cyclin E-CDK2 activation and the increase in p27 accumulation via blockade of the PDGFR^-ERK1/2 signaling cascade.

Activation of JAK-STAT pathway is required for platelet-derived growth factor-induced proliferation of pancreatic stellate cells

AM: To clarify the role of Janus kinase-signal transducers and activators of transcription (JAK-STAT) pathway in platelet-derived growth factor (PDGF) induced proliferation in activated pancreatic stellate cells (PSCs). METHODS: PSCs were isolated from rat pancreas tissue, and used in their culture-activated, myofibroblast-like phenotype. STAT-specific binding activity was assessed by electrophoretic mobility shift assay. Activation of Src, JAK2, STAT1, STAT3, and ERK was determined by Western blotting using anti-phosphospecific antibodies. Cell proliferation was assessed by measuring the incorporation of 5-bromo-2'-deoxyuridine. RESULTS: PDGF-BB induced STAT-specific binding activity, and activation of Src, JAK2, STAT1, STAB, and ERK. Ethanol and acetaldehyde at clinically relevant concentrations decreased basal activation of JAK2 and STAT3. PDGF-induced activation of STAT1 and STAT3 was inhibited by a Src inhibitor PP1 and a JAK2 inhibitor AG490, whereas PDGF-induced activation of ERK was inhibited by PP1, and not by AG490. PDGF-induced proliferation was inhibited by PP1 and AG490 as well as by STAT3 antisense oligonucleotide. CONCLUSION: PDGF-BB activated JAK2-STAT pathway via Src-dependent mechanism. Activation of 3AK2-STAT3 pathway, in addition to ERK, may play a role in PDGF-induced proliferation of PSCs.

S100A6 gene have a positive influence on the growth and proliferation of gastric cancer cell MKN45

Objective： S100A6 （a.k.a., calcyclin） is over-expressed in several human tumors, including gastric carcinoma, human melanoma, pancreatic carcinoma, squamous cell carcinoma, malignant fibrous histiocytoma （MFH）, and carcinomas of the thyroid, breast, and colon. However, little is known about the role S100A6 plays in gastric adenocarcinoma. In the present study, we intended to investigate the influence of S100A6 on the growth, proliferation, apoptosis, invasion and cell cycle of the gastric cancer cell MKN45. Methods： As an important member of S100 family, S100A6 cDNAwas subcloned into a constitutive vector pcDNA3.1 followed by transfection in gastric cancer cell line MKN45 by using liposome. Then stable transfectants were selected and appraised. The apoptosis and cell cycles of these clones were analyzed by using flow cytometric assay. The growth and proliferation were analyzed by cell growth curves and colony-forming assay respectively. The S100A6 stable expression clones （MKN-S100A6） were detected and compared with their control groups respectively. Results： MKN- S100A6 grew faster than MKN45 and MKN-PC （MKN45 transfected with pcDNA3.1 vector）. The cell counts of MKN-SI00A6 in the fifth, sixth and seventh days were significantly more than those of control groups （P 〈 0.05）. Cell cycle analysis showed that proportions of MKN-S100A6 in G0-G1 and G2-M were different significantly with those of its control groups respectively （P 〈 0.05）. The apoptosis rate of MKN-S100A6 was significantly lower than those of control groups （P 〈 0.05）. Results of colony-forming assay showed that the colon formation rate of MKN-S100A6 was higher than those of control groups （P 〈 0.05）. Conclusion： S100A6 can promote the growth and proliferation of gastric cancer cells. It can help tumor cell maintain malignant phenotype. In gastric cancer, S100A6 could be thought as a tumor-enhancing gene in some distance, but its role could be complicated.

Effect of Spatholobus suberectus Dunn on proliferation of progenitor cells in mice with bone marrow depression

AIM： To study the effect of Spatholobus suberectus Dunn on the prolilferation and hematonic mechanism of Spatholobus suberectus Dunn. Methods： The techniques of culture of hematopoietic cell and hematopoietic growth factor （HGF） assay were used. The method of semi-solid culture with methylcellulose of CFU-GM, CFU-E, BFU-E, CFU-Meg was adopted in bone marrow depressed mice which treated with Spatholobus suberectus Dunn for a long time. Results： Spatholobus suberectus Dunn could obviously promote the proliferation of bone morrow cells and spleen lymphocytes in healthy and anaemic mice. The cuhure medium of spleen cell, macrophage, lung and skeletal muscle treated with Spatholobus suberectus Dunn had much stronger stimulating effects on hematopoietic cells. The numbers of CFU-GM, CFU-E, BFU-E, CFU-Meg in bone marrow depressed mice were raised distinctly under the control of Spatholobus suberectus Dunn as compared with those of contrast group. Conclusions： Spatholobus suberectus Dunn may enhance hematopoiesis by stimulating directly and/or indirectly stroma cell in hematopoietic inductive microenvironment and muscle tissue to secrete some HGF （Epo, GM-CSF, IL, and MK-CSF）. It can promote the proliferation and differentiation of hematopoietic cells in anaemic mice. This is one of the biological mechanisms for hematonic effect of Spatholobus suberectus Dunn.

Effects on Proliferation and Migration of the Human Colon Carcinoma Cell Line SW620 by Silencing of Hepatocyte Growth Factor Expression

OBJECTIVE Hepatocyte growth factor （HGF） expression is closely related to the progression and poor prognosis of colorectal cancer patients. In this study, we investigated the effects on proliferation and migration of the human colon carcinoma cell line SW620 by silencing HGF expression. METHODS HGF was silenced using specific HGF a/f3 siRNA. The proliferation, migration, cell cycle and ultrastructure of SW620 cells were examined. RESULTS The transfection efficiency was 70%-80%. The expression rate of HGF in the experimental group was significantly lower than that in the negative and blank control groups （P 〈 0.05）. The proliferation inhibition rate in the experimental group at 24, 48, 72 and 96 h after transfection was 14.2%, 50.2%, 39.5% and 23.2%, respectively. The migratory ability of cells in the experimental group was significantly inhibited compared with that in the negative control or blank control groups （58.2% vs. 2.1% or 0%, P 〈 0.05）. CONCLUSION The application of RNA interference to silence the expression of HGF in the colon carcinoma cell line SW620 effectively inhibits the proliferation and migration of tumor cells.

Involvement of 90-kuD ribosomal S6 kinase in collagen type Ⅰ expression in rat hepatic fibrosis

AIM： To investigate the relationship between 90-kuD ribosomal $6 kinase （pg0RSK） and collagen type I expression during the development of hepatic fibrosis in vivo and in vitro.METHODS： Rat hepatic fibrosis was induced by intraperitoneal injection of dimethylnitrosamine. The protein expression and cell location of p90RSK and their relationship with collagen type I were determined by co-immunofluoresence and confocal microscopy.Subsequently, RNAi strategy was employed to silence p90RSK mRNA expression in HSC-T6, an activated hepatic stellate cell （HSC） line. The expression of collagen type I in HSC-T6 cells was assessed by Western blotting and real-time polymerase chainreaction. Furthermore, HSCs were transfected with expression vectors or RNAi constructs of p90RSK to increase or decrease the p90RSK expression, thencollagen type I promoter activity in the transfected HSCs was examined by reporter assay. Lastly HSC-T6 cells transfected with p90RSK siRNA was treated withor without platelet-derived growth factor （PDGF）-BB at a final concentration of 20μg/L and the cell growthwas determined by MTS conversion.RESULTS： In fibrotic liver tissues, p90RSK was over-expressed in activated HSCs and had a significantpositive correlation with collagen type I levels.In HSC-T6 cells transfected with RNAi targeted top90RSK, the expression of collagen type I was down-regulated （61.8% in mRNA, P 〈 0.01, 89.1% inprotein, P 〈 0.01）. However, collagen type ] promoteractivity was not increased with over-expression of p90RSK and not decreased with low expression either,compared with controls in the same cell line （P = 0.076）.Furthermore, p90RSK siRNA exerted the inhibitionof HSC proliferation, and also abolished the effect of PDGF on the HSC proliferation.CONCLUSION： p90RSK is over-expressed in activatedHSCs and involved in regulating the abnormalexpression of collagen type I through initiating theproliferation of HSCs.

Morphological Evaluation of Adhesion and Proliferation of Osteoblast Like Cells Grown on Gelatin/Genipin Scaffold

This work focusing on studying the biocompatibility and the effect of gelatin porous scaffold on the characteristics of human osteoblast like cells, including proliferation, adhesion, scaffold-cell interaction and its potential to induce bone regeneration. Osteoblast like cells were seeded on gelatin/genipin scaffolds for 7, 14 and 21 days. Cell proliferation assay, light microscopy, transmission electron microscopy and high resolution scanning electron microscopy were carried to evaluate cell viability, cell adhesion and the production of extracellular matrix. Cell proliferation assay showed a high biocompatibility of the material. High resolution scanning electron microscopy and light microscopy showed a strong adhesion of MG63 ceils on the surface of gelatin scaffold and high penetration in the macroporosities of the material. TEM analysis showed an intense production of extracellular matrix protein. In vitro analysis indicated a good biocompatibility of the scaffold and presents it as a potential candidate material for tissue engineering.

In vitro and in vivo suppression of hepatocellular carcinoma growth by midkine-antisense oligonucleotide-loaded nanoparticles

AIM:To synthesize antisense oligonucleotides (ASODNs) of midkine (MK), package the ASODNs with nanoparticles, and to inhibit hepatocellular carcinoma (HCC) growth using these nanoparticles.METHODS: HepG2 cell proliferation was analyzed in vitro using the 3-(4,5-dimethythiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2Htetrazolium, inner salt assay. The in vivo activity of nanoparticles delivering the MK-ASODNs was analyzed by histopathological and immunohistochemical staining and quantitative real time polymerase chain reaction (PCR). RESULTS: The in vitro proliferation of HepG2 cells was signif icantly inhibited by the nanoparticles packaged with MK-ASODNs (NANO-ASODNs). Furthermore, the NANO- ASODNs signif icantly inhibited the growth of HCC in the mouse model. CONCLUSION: NANO-ASODNs can significantly suppress the growth of HCC in vitro and in vivo.