珍珠层人工骨对成骨细胞在体内外增殖的作用

探讨珍珠层／聚乳酸人工骨的生物相容性，为其作为生物源性成骨材料和组织工程骨支架材料提供可行性依据。本实验进行珍珠层／聚乳酸人工骨复合成骨细胞后在体内外增殖的研究。

CD40调节胰腺癌细胞生物学行为的研究

目的:研究CD40L对胰腺癌细胞生物学特性、干性的影响。方法:CCK-8活细胞计数法分析CD40可溶性配体(s CD40L)不同浓度(0、1、2、4μg/m L)在不同时间对人胰腺癌细胞系panc02生长增殖的影响,Annexin V/PI双染法检测凋亡;细胞划痕实验检测迁移的改变。Q-PCR检测4μg/m L s CD40L处理panc02 48 h后c-Myc、OCT-4、Sox-2水平的改变。建立Balb/c裸鼠胰腺癌荷瘤模型,随机分为2组,对照组(生理盐水)和s CD40L(10 mg/kg)组,分别腹腔注射s CD40L 10mg/kg以及等体积生理盐水,每天注射3次,游标卡尺测量0、7、14、21、28 d时肿瘤体积。结果:与对照组比较,浓度为1、2、4μg/m L的s CD40L处理panc02 96 h时可显著抑制panc02的增殖(P<0.01),并促进panc02的凋亡(P<0.01),并呈剂量依赖性地抑制panc02的迁移能力(P<0.01);s CD40L 4μg/m L组可明显抑制panc02细胞的cMyc、OCT-4、Sox-2的表达(P<0.01)。在裸鼠荷瘤实验中,与对照组比较,s CD40L处理28 d明显抑制肿瘤体积(P<0.01,252±13 vs.189±9)。结论:CD40通路的活化能够抑制胰腺癌细胞增殖迁移、促进凋亡,同时抑制胰腺癌细胞系体内的增殖能力,而这一效应可能与CD40L抑制胰腺癌细胞系干性相关。

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.

Octreotide inhibits proliferation and invasion of MHCC97-H cells in vitro and in vivo

Objective: To figure out the effect of somatostatin analogue Octreotide on proliferation and invasion of human hepatocellular carcinoma cell MHCC97-H and the underlying mechanism in vitro and in vivo. Methods: MHCC97-H cells were treated with Octreotide at the concentration of 0.2 ug/mL in vitro, proliferation related to time was evaluated. After treated with Octreotide at the concentration of 0.2 ug/mL for 48 h, MHCC97-H cells were observed by transmission electron microscope. Cell proliferation was detected by MTT assay after MHCC97-H cells were treated with Octreotide at different concentrations including 0.05, 0.1, 0.2, 0.4, 0.6 and 0.8 ug/mL for 36 h in vitro. 27 nude mice, in which MHCC97-H tumor mass was planted orthotopically, were divided into 3 groups randomly including control group (intraperitoneal injection with equal volume normal saline; n=8), low dose treated group (intraperitoneal injection with Octreotide at 50 ug/kg?d; n=9) and large dose treated group (intraperitoneal injection with Octreotide at 200 ug/kg?d; n=10). All mice were raised for 35 d and sacrificed. The information about survival time, the weight at death point and the pathology change of liver and lung was collected. The expression of vascular endothelial growth factor (VEGF) and matrix metalloproteinases-2 (MMP-2) in mouse HCC tissues were detected by immunohistochemistry finally. Results: MTT assays showed that Octreotide inhibited the proliferation of MHCC97-H cells significantly. Apoptosis cells were found by transmission electron microscope after treatment with Octreotide at 0.2 ug/mL for 48 h in vitro. The proliferation was inhibited significantly by Octreotide in a dose-dependant manner (r=0.86, P<0.01). Compared with control group, the treated group had the heavier weight at death point and lower intrahepatic metastasis ratio (P<0.05), meanwhile, there was not significant difference in treated groups (P>0.05). The positive expression ratios of VEGF and MMP-2 in treated groups were lower than those in control group (P<0.05), while there was no apparent difference in treated groups (P>0.05). Conclusion: Octreotide could inhibit the proliferation of MHCC97-H cells in vitro via inducing apoptosis and the inhibitory function acts in a dose-dependant manner. Octreotide could improve survival of mice with MHCC97-H cells and inhibit the metastasis of MHCC97-H cells in vivo. Regulation of VEGF and MMP-2 expression by Octreotide would be involved in its inhibition in vivo.

Anti-angiogenesis effect of Demethyl bryoanthrathiophene(DBT) in vitro

Objective:The aim of the study was to investigate the effect of Demethyl bryoanthrathiophene(DBT) on proliferations of human umbilical vein endothelial cells(HUVECs) and human lung adenocarcinoma cell line A549,and antiangiogenic effect of DBT on HUVECs in vitro.Methods:MTT assay was used to observe the effect of DBT on proliferations of HUVECs and A549 cells,flat plate scarification assay and tube formation in vitro test were used to observe the impact of DBT on migration and vaso-formed ability of HUVECs.The effects of DBT on apoptosis and cell cycle of HUVECs were calculated by flow cytometry.Results:MTT assay showed that treatment with DBT resulted in strong inhibition to the growth of HUVECs and A549 cells.The inhibition effects of DBT on HUVECs and A549 cells were related to dosage and times of dependency.In different doses of DBT(0.16,0.32 and 0.48 μmol/L) of flat plate scarification for 24 h,inhibition rates of DBT to migration of HUVECs were 14.70%,38.23% and 58.82%,respectively.In dose of DBT from 0.04,0.20 to 0.40 μmol/L for 24 h in tube formation,there were significance differences(P < 0.01) in the decreasing number of angiogenesis and incomplete blood vessel compared with control groups.All results showed that DBT promoted the apoptosis rate of HUVECs,and the increase of concentration of DBT accompanied the acceleration of apoptosis rate.Conclusion:DBT could inhibit the proliferations of HUVECs and A549 cells,and effectively suppress angiogenesis in vitro.

ADAMTS-7 promotes vascular smooth muscle cells proliferation in vitro and in vivo

Vascular smooth muscle cell(VSMC) proliferation and migration are pivotal for the pathogenesis of atherosclerosis and post-angioplasty restenosis. We have recently reported that a disintegrin and metalloproteinase with thrombospondin motifs-7(ADAMTS-7), a novel metalloproteinase, contributes directly to neointima formation by mediating VSMC migration. However, whether ADAMTS-7 affects VSMC proliferation remains unclear. In this study, we found that luminal adenoviral delivery of ADAMTS-7 aggravated intimal hyperplasia 7 d after injury, paralleled by an increased percentage of PCNA-positive cells in both intima and media. In contrast, perivascular administration of ADAMTS-7 si RNA, but not scrambled si RNA to injured arteries attenuated intimal thickening at day 7, paralleled with reduced intimal VSMC replication, without alteration of VSMC proliferation in the media. In accordance, [3H]-thymidine incorporation assay in primary cultured rat VSMCs revealed an enhanced replication rate(by 61%) upon ADAMTS-7 overexpression and retarded proliferation(by 23%) upon ADAMTS-7 si RNA administration. Our data demonstrates that ADAMTS-7 promotes VSMC proliferation both in vitro and in vivo. ADAMTS-7 may therefore serve as a novel therapeutic target for atherosclerosis and post-angioplasty restenosis.