IFN、IL—6联合丝裂霉素抑制肺腺癌A549细胞系生长的研究

肺癌是呼吸系统恶性肿瘤之一,近年来其发病率、死亡率急剧上升。干扰素(IFN)及白介素6(IL—6)是细胞因子网络中的重要组成部分,具有广泛的生物活性,在体内外都具有较强的抗肿瘤作用。丝裂霉素C(MMC)是临床上常用的抗肿瘤药物,能抑制肿瘤细胞DNA的复制而发挥其抗肿瘤作用。

基因重组人生长激素对人肝癌细胞系生长的影响

目的 研究基因重组人生长激素 (rhGH )作用人肝癌细胞系后细胞周期动力学的改变 ,了解其对人肝癌细胞系生长的影响。方法 不同浓度rhGH ( 0、2、5、2 5、5 0、15 0、5 0 0 μg/L)作用于人肝癌细胞系HepG2 ,采用流式细胞仪检测作用 2 4、48、72h后的细胞周期动力学 ,分析其增殖指数、S期细胞百分比及细胞凋亡率的变化。结果 rhGH作用后的HepG2细胞周期动力学改变与作用时间无显著相关 ;与对照组比较 ,在生理浓度 ( 2 5 μg/L)rhGH作用下 ,PI及S %显著上升(P <0 .0 5 ) ,但在超生理浓度 ( 5 0 0 μg/L)rhGH作用下 ,PI及S %无显著改变 ( P >0 .0 5 ) ;另 2、5、2 5、5 0、15 0、5 0 0 μg/LrhGH均诱导人肝癌细胞凋亡 ,且呈浓度 效应关系。 结论 在体外培养体系中 ,rhGH在低于或等于生理浓度时 ,促进人肝癌细胞增殖 ;在高于生理浓度时 ,抑制人肝癌细胞增殖。

胞嘧啶脱胺酶/5-氧胞嘧啶基因系统对前列腺癌细胞系生长的抑制作用

目的 探讨胞嘧啶脱胺酶 (cytosinedeaminase,CD) / 5 氟胞嘧啶基因 (5 FC)基因系统对体外前列腺癌细胞生长的作用。 方法 经基因重组并纯化的腺病毒AdCMVCD分别感染体外前列腺癌细胞DU 14 5、RM 1,观察CD/ 5 FC系统对细胞的生长抑制作用和旁观者效应。 结果 CD/5 FC系统能明显抑制体外前列腺癌细胞系DU14 5、RM 1的生长 ,经 10 0m .o .i(multiplicityofinfec tion)AdCMVCD感染并加入 15 .5mmol/L 5 FC后 4、6d ,细胞抑制率高于 97% ;同时此系统具有很强的旁观者效应 ,当受染细胞混合比例为 10 %时 ,即有 >75 %的细胞被杀死。 结论 CD/ 5 FC系统是一种有效的基因治疗方法 ,旁观者效应为此系统在实体肿瘤中的应用提供了可行性。

非生长因子依赖的MDS细胞系的建立及其生物学特性研究

本研究的目的是从骨髓增生异常综合征-慢性粒-单核细胞白血病(MDS-CMML)病人获取骨髓细胞,建立 非生长因子依赖的细胞株。该细胞株在不添加任何生长因子的含15％胎牛血清的RPMI 1640和DMEM混合培养 液中进行了培养,并分别从细胞株的形态、细胞表面抗原分子、细胞增殖、分化、凋亡等方面检测其生物学特性。结 果表明:在不添加任何生长因子的培养条件下该细胞株可以长期存活和生长,并能向单核及巨核细胞分化。结论: 成功建立了一株非生长因子依赖的MDS-JSN04(MDS-江苏南京04)细胞株,并阐明了其部分生物学特性。

粒细胞巨噬细胞集落刺激因子对人卵巢癌细胞株生长的影响

目的：了解粒细胞巨噬细胞集落刺激因子（ＧＭＣＳＦ）对人卵巢癌细胞株生长的影响。方法：采用ＸＴＴ比色法及流式细胞仪检测ＧＭＣＳＦ对卵巢癌细胞株ＳＫＯＶ３、３ＡＯ及ＣＡＯＶ３ 细胞生长的影响。结果：用ＸＴＴ法检测ＧＭＣＳＦ对ＳＫＯＶ３、３ＡＯ 及ＣＡＯＶ３ 细胞的生长均无明显影响；流式细胞仪检测也未发现ＧＭＣＳＦ对ＳＫＯＶ３、３ＡＯ及ＣＡＯＶ３细胞的增殖指数有明显影响。结论：在卵巢癌化疗过程中应用ＧＭＣＳＦ是比较安全的，对其潜在的促进肿瘤细胞增殖的顾虑也许是不必要的。

胶质细胞系源性神经营养因子对泻药性结肠大鼠肠道传输功能的影响

背景:慢传输型便秘(STC)患者长期依赖泻剂排便,导致泻药性结肠,部分患者最终需手术切除结肠。目的:探讨胶质细胞系源性神经营养因子(GDNF)对泻药性结肠大鼠肠道传输功能的影响。方法:以大黄灌胃建立大鼠STC模型。将大鼠随机分为正常NaCl组、正常GDNF组、模型NaCl组和模型GDNF组。正常GDNF组和模型GDNF组大鼠尾静脉注射GDNF,其余两组注射0.9%NaCl溶液。1周后处死大鼠,以墨汁推进试验测定肠道传输功能,并行结肠组织HE染色。结果:模型NaCl组肠道推进率显著低于正常NaCl组(P<0.01);模型GDNF组推进率显著高于模型NaCl组(P<0.01),结肠黏膜组织学表现较模型NaCl组有所改善。结论:外源性GDNF可明显改善大鼠肠道传输功能。

hsa-mir-615过表达胰腺癌稳定细胞亚系的构建及功能初步验证

慢病毒（Lentivirus）载体可以将外源基因有效地整合到宿主染色体上,从而达到持久性表达[1].本实验在对胰腺癌差异表达的microRNA进行研究时发现hsa-mir-615在胰腺癌组织和胰腺正常导管上皮组织中呈现差异表达[2],异常表达的microRNA可能具有生物学功能.因此,本实验构建hsa-mir-615过表达慢病毒载体并包装成慢病毒,感染人胰腺癌细胞MIA PaCa-2,建立hsa-mir-615过表达稳定细胞系,使用CCK-8法观察hsa-mir-615对胰腺癌细胞系生长的影响,为进一步研究hsa-mir-615的生物功能构建细胞模型.

1950 MHz GSM-Talk信号对睾丸支持细胞增殖和分泌功能的影响

采用贴壁生长的小鼠睾丸支持细胞(TM4)在含10%胎牛血清的DMEM/F12 1:1培养液中培养至指数生长期后,消化制备成1.5×103个·m L-1的细胞悬液,接种于35 mm培养皿中,每皿接种3 m L,接种12个皿。细胞接种后24 h更换新鲜培养液,然后将培养皿随机分为假辐照组和辐照组,每组6个皿。将2组细胞分别置于辐照装置的2个小室中,其中一个小室产生GSM-Talk信号,1950 MHz连续波,比吸收率为3 W·kg-1,辐照时间为5 d;另一个小室不产生GSM信号,用于假辐照。辐照结束后1-5 d,采用CCK-8和Brd U法检测细胞的增殖情况,免疫荧光染色检测细胞增殖标记物Ki67的蛋白表达,ELISA法检测小鼠干细胞因子(SCF)和小鼠胶质细胞系来源的神经生长因子(GDNF)的水平。结果显示:与对照组相比,辐照组TM4细胞的增殖能力明显减弱(p<0.05),细胞增殖标记物Ki67的表达亦有减弱趋势,细胞上清中SCF水平明显降低(p<0.05),而GDNF水平明显升高(p<0.05)。实验结果提示,GSM-Talk连续辐照5 d可抑制TM4细胞的增殖并影响其分泌功能。

Effects of basic fibroblast growth factor on ischemic gut and liver injuries

AIMS To explore the possible effects of basic fibrob- last growth factor (bFGF) on ischemic gut and liver in- juries after trauma. METHODS Animal model of super mesenteric artery (SMA) occlusion was used in this study. Seventy-two Wistar rats were divided into three groups of 24 rats each. Each animal in group 1 (bFGF treated) was in- jected with 4 μg of bFGF in 0.15 ml of normal saline solution containing 0.1%(w/v) heparin through the jugular vein at the onset of reperfusion. Animals in group 2 (saline treated) received the same vehicle, but without bFGF. Group 3 (sham-operated) ani- mals were treated with the same operations,but without SMA occlusion. Liver function parameters, serum TNFα,bacterial examination and pathological study were used to evaluate the results. RESULTS In group 1,the amounts of ALT and AST and serum TNFα were reduced significantly at 6,24 and 48 hours as compared with group 2. Bacterial ex- amination showed that the bacterial translocation from gut to liver,spleen and MLN in group 1 was much lower than that in group 2. The pathological results support the concept of significant protecting effects of bFGF. CONCLUSIONS Venous administration of bFGF may help reduce gut and liver injuries after ischemia and reperfusion. Its mechanism of action may involve the mitogenic and non-mitogenic effects of bFGF.

胃、十二指肠

P物质对胃癌细胞增殖的作用;胃肠道间质瘤196例临床分析;胃窦癌肝十二指肠韧带淋巴结微转移与清扫;三孔法腹腔镜胃十二指肠溃疡穿孔修补术;丁酸钠对胃癌细胞系生长的抑制作用及其机制;

颅脑

热休克对人胶质瘤热休克蛋白70表达水平的影响；重组腺病毒携带外源P16基因抑制人脑恶性胶质瘤细胞系生长的研究；硼中子浮获法诱导C6胶质瘤细胞凋亡；MDR1小干扰RNA调节人脑多形性胶质母细胞瘤细胞系BT325的药物敏感性;56例瘤周脑水肿与脑膜瘤增殖及术后复发的临床意义；急性垂体腺瘤卒中病人行经蝶手术前后的垂体功能；

The promoting molecular mechanism of alphafetoprotein on the growth of human hepatoma Bel7402 cell line

AIM: The goal of this study was to characterize the AFP receptor, its possible signal transduction pathway and its proliferative functions in human hepatoma cell line Bel 7402. METHODS: Cell proliferation enhanced by AFP was detected by MTT assay, 3H-thymidine incorporation and S-stage percentage of cell cycle analysis. With radioactive labeled 125I-AFP for receptor binding assay; cAMP accumulation, protein kinase A activity were detected by radioactive immunosorbent assay and the change of intracellular free calcium (Ca2+i) was monitored by scanning fluorescence intensity under TCS-NT confocal microscope. The expression of oncogenes N- ras, p 53, and p21( ras ) in the cultured cells in vitro were detected by Northern blotting and Western blotting respectively. RESULTS: It was demonstrated that AFP enhanced the proliferation of human hepatoma Bel 7402 cell in a dose dependent fashion as shown in MTT assay, (3)H-thymidine incorporation and S-phase percentage up to 2-fold. Two subtypes of AFP receptors were identified in the cells with Kds of 1.3 x 10(-9)mol.L(-1) and 9.9 x10(-8)mol. (-1)L respectively. Pretreatment of cells with AFP resulted in a significant increase (625%) in cAMP accumulation. The activity of protein kinase A activity were increased up to 37.5, 122.6, 73.7 and 61.2% at treatment time point 2, 6, 12 and 24 hours. The level of intracellular calcium were elevated after the treatment of alpha-fetoprotein and achieved to 204% at 4 min. The results also showed that AFP(20mg.L(-1)) could upregulate the expression of N- ras oncogenes and p 53 and p21( ras ) in Bel 7402 cells. In the later case,the alteration were 81.1%(12h) and 97.3%(12h) respectively compared with control. CONCLUSION: These results demonstrate that AFP is a potential growth factor to promote the proliferation of human hepatoma Bel 7402 cells. Its growth-regulatory effects are mediated by its specific plasma membrane receptors coupled with its transmembrane signaling transduction through the pathway of cAMP-PKA and intracellular calcium to regulate the expression of oncogenes.

The Inhibitory Effect of Oridonin on the Growth of Fifteen Human Cancer Cell Lines

OBJECTIVE To study the inhibitory effect of oridonin on the growth of cancer cells. METHODS Fifteen human cancer cell lines were subjected to various concentrations of oridonin in culture medium. The inhibitory rate of cell growth was measured by the MTT assay, and compared with a negative control and 5-Fu-positive control. RESULTS The 50% inhibiting concentration （IC50） and maximal inhibition （Imax） of oridonin shown by studying the growth of the cancer cell lines were as follows： leukemias （HL60 cells： 3.9 μg/ml and 73.8%, K562 cells： 4.3 μg/ml and 76.2%）; esophageal cancers（SHEEC cells： 15.4 μg/ml and 99.2%, Eca109 cells： 15.1 μg/ml and 84.6%, TEl cells： 4.0 μg/ml and 70.2%）; gastric cancers （BGC823 cells： 7.6 μg/ml and 98.7%, SGC7901 cells： 12.3 μg/ml and 85.7%）; colon cancers （HT29 cells： 13.6 μg/ml and 97.2%, HCT cells： 14.5 μg/ml and 96.5%）; liver cancers （Bel7402 cells： 15.2 μg/ml and 89.2%, HepG2 cells： 7.1 μg/ml and 88.3%）; pancreatic cancer （PC3 cells： 11.3 μg/ml and 68.4%）; lung cancer （A549 cells： 18.6 μg/ml and 98.0% ）; breast cancer （MCF7 cells： 18.4 μg/ml and 84.7%）; uterine cervix cancer （Hela cells： 13.7 μg/ml and 98.5%）. CONCLUSION Oridonin had a relatively wide anti-tumor spectrum, and a relatively strong inhibitory effect on the growth of the 15 human cancer cells. Inhibitory effects were concentration dependent.

Genomic imbalances in esophageal carcinoma cell lines involve Wnt pathway genes

AIM： To identify molecular markers shared across South African esophageal squamous cell carcinoma （ESCC） cell lines using o/togenetics, fluorescence in situ hybridization （FISH） and single nucleotide polymorphism （SNP） array copy number analysis. METHODS： We used conventional cytogenetics, FISH, and multicolor FISH to characterize the chromosomal rearrangements of five ESCC cell lines established in South Africa. The whole genome copy number profile was established from 250K SNP arrays, and data was analyzed with the CNAT 4.0 and GISTIC software. tions involved the following chromosomal regions and genes： 11q13.3 （CCND1, FGF3, FGF4, FGF19, MYEOV）, 8q24.21（C-MYC, FAM84B）, 11q22.1-q22.3 （B[RC2, BIRC3）, 5p15.2 （CTNND2）, 3qll.2-q12.2 （MINA） and 18p11.32 （TYMS, YES1）. The significant deletions included 1p31.2-p31.1 （CTH, GADD45a, DIRAS3）, 2q22.1 （LRPIB）, 3p12.1-p14.2 （FHIT）, 4q22.1-q32.1 （CASP6, SMAD1）, 8p23.2-q11.1 （BNIP3L） and 18q21.1-q21.2 （SMAD4, DCC）. The 3p11.2 translocation breakpoint was shared across four cell lines, supporting a role for genes involved at this site, in particular, the EPHA3 gene which has previously been reported to be deleted in ESCC.CONCLUSION： The finding that a significant number of genes that were amplified （FGF3, FGF4, FGF19, CCND1 and C-MYC） or deleted （SFRP2 gene） are involved in the Wnt and fibroblast growth factor signaling pathways, suggests that these pathways may be activated in these cell lines.

An R2R3-type transcription factor gene AtMYB59 regulates root growth and cell cycle progression in Arabidopsis

MYB proteins play important roles in eukaryotic organisms. In plants, the R1R2R3-type MYB proteins function in cell cycle control. However, whether the R2R3-type MYB protein is also involved in the cell division process remains unknown. Here, we report that an R2R3-type transcription factor gene, AtMYB59, is involved in the regulation of cell cycle progression and root growth. The AtMYB59 protein is localized in the nuclei of onion epidermal cells and has transactivation activity. Expression of AtMYB59 in yeast cells suppresses cell proliferation, and the transfor- mants have more nuclei and higher anenpioid DNA content with longer cells. Mutation in the conserved domain of AtMYB59 abolishes its effects on yeast cell growth. In synchronized Arabidopsis cell suspensions, the AtMYB59 gene is specifically expressed in the S phase during cell cycle progression. Expression and promoter-GUS analysis reveals that the AtMYB59 gene is abundantly expressed in roots. Transgenic plants overexpressing AtMYB59 have shorter roots compared with wild-type plants （Arabidopsis accession Col-0）, and around half of the mitotic cells in root tips are at metaphase. Conversely, the null mutant myb59-1 has longer roots and fewer mitotic cells at metaphase than Col, suggesting that AtMYB59 may inhibit root growth by extending the metaphase of mitotic cells. AtMYB59 regulates many downstream genes, including the CYCB1;1 gene, probably through binding to MYB-responsive elements. These results support a role forAtMYB59 in cell cycle regulation and plant root growth.

INHIBITORY EFFECTS OF MIFEPRISTONE ON THE GROWTH OF HUMAN GASTRIC CANCER CELL LINE MKN-45 IN VITRO AND IN VIVO

Objective To explore the effects of mifepristone on the growth of human gastric cancer cell line MKN-45 and its possible mechanisms. Methods In situ hybridization was used to detect the expression of progesterone receptor (PR) mRNA in MKN-45 cells. Proliferation, cell cycle distribution, and the expression of Bcl-xL and vascular endothelial growth factor (VEGF) of MKN-45 cells incubated with various concentrations of mifepristone (1, 5, 10, and 20 μmol/L) were analyzed using MTT reduction assay, flow cytometry, reverse transcription-polymerase chain reaction (RT-PCR), and enzyme-linked immunoab-sorbent assay (ELISA), respectively. After transplantation of MKN-45 cells underneath the skin of athymic mice, mifepristone was administrated with the dose of 50 mg/(kg·d) for 6 weeks to evaluate the tumor growth. Apoptosis and the expression of proliferating cell nuclear antigen (PCNA) in xenografted tumors were detected using transmission electron microscopy and immunohistochemical staining, respectively. Results PR mRNA was highly expressed in cultured MKN-45 cell. Mifepristone dose-dependently inhibited the pr-oliferation of MKN-45 cells, and the inhibitory rate was dramatically increased from 7.21% to 47.23%. The inhibitory effect was accompanied by a dose-dependent increase in the percentage of cells in G 0 /G 1 phase, and with a concurrent decrease in the proportion of S- and G 2 /M-phase cells and the proliferative index from 57.65% to 24.54%. Meanwhile, mifepristone down-regulated the expression of Bcl-xL and VEGF in a dose-dependent manner. In vivo, mifepristone effectively inhibited the growth of xenografted tumors in nude mice (55.14% for inhibitory rate), induced apoptosis, and down-regulated PCNA expression in gastric cancer. Conclusion Mifepristone exerts significant growth inhibitory effects on PR-positive human MKN-45 gastric cancer cells via multiple mechanisms, and may be a beneficial agent against the tumor.

Growth inhibitory effect of wild-type Kras2 gene on a colonic adenocarcinoma cell line

AIM： To observe the growth inhibitory effect of wild-type Kras2 gene on a colonic adenocarcinoma cell line Caco-2. METHODS： Recombinant plasmid pCI-neo-Kras2 with wild type Kras2 open reading frame was constructed. The Caco-2 cells were transfected with either pCI-neo or pCI-neo-Kras2 using Upofectamine 2000. The expression of wild type Kras2 was examined by Northern blot analysis. And the expression of wild type Kras2 protein was examined by Western blot analysis. The effects of wild-type Kras2 on cell proliferation were analyzed by monotetrazolium （MTT） assay, meanwhile analyses of cell cycle and spontaneous apoptosis rate were carried out by flow cytometry （FCM）. RESULTS： The plasmid of pCI-neo-Kras2 was successfully established. The growth rate of cells transfected with pCI-neo-Kras2 was significantly lower than the control cells transfected with the empty pCI- neo vector （P 〈 0.05）. Cell cycle analysis revealed arrest of the pCI-neo-Kras2 transfected cells in G0/G1 phases, decreased DNA synthesis and decreased fractions of cells in S phase. The proliferative index of cells transfected with pCI-neo-Kras2 was decreased compared with the control cells （49.78% vs 64.21%）, while the apoptotic rate of Caco-2 cells with stable Kras2 expression increased （0.30% vs 0.02%）. CONCLUSION： The wild-type Kras2 gene effectively inhibits the growth of the colonic adenocarcinoma cell line Caco-2.

Renin-angiotensin system in the pathogenesis of liver fibrosis

Hepatic fibrosis is considered a common response to many chronic hepatic injuries. It is a multifunctional process that involves several cell types, cytokines, chemokines and growth factors leading to a disruption of homeostatic mechanisms that maintain the liver ecosystem. In spite of many studies regarding the development of fibrosis, the understanding of the pathogenesis remains obscure. The hepatic tissue remodeling process is highly complex, resulting from the balance between collagen degradation and synthesis. Among the many mediators that take part in this process, the components of the Renin angiotensin system （RAS） have progressively assumed an important role. Angiotensin （Ang） II acts as a profibrotic mediator and Ang-（1-7）, the newly recognized RAS component, appears to exert a counter-regulatory role in liver tissue. We briefly review the liver fibrosis process and current aspects of the RAS. This review also aims to discuss some experimental evidence regarding the participation of RAS mediators in the pathogenesis of liver fibrosis, focusing on the putative role of the ACE2-Ang-（1-7）- Mas receptor axis.

Gfi1.1 regulates hematopoietic lineage differentiation during zebrafish embryogenesis

Growth factor independence 1 （GFI1） is important for maturation of mammalian lymphocytes and neutrophils and maintenance of adult hematopoietic stem cells （HSCs）. The role of GFI1 in embryonic hematopoiesis is less well characterized. Through an enhancer trap screen and bioinformatics analysis, we identified a zebrafish homolog of Gill （named grill） and analyzed its function during embryonic development. Expression of both an endogenous griLl gene and a GFP reporter gene inserted near its genomic locus was detected in hematopoietic cells of zebrafish embryos. Morpholino （MO） knockdown of gill.1 reduced expression of scl, Imo2, c-myb, mpo, ragl, gatal and hemoglobin alpha embryonic-1 （hbael）, as well as the total amount of embryonic hemoglobin, but increased expression ofpu.1 and l-plastin. Under the same conditions, MO injection did not affect the markers involved in vascular and pronephric development. Conversely, overexpression of gill.1 via mRNA injection enhanced expression ofgatal but inhibited expression ofpu.1. These findings suggest that Gill.1 plays a critical role in regulating the balance of embryonic erythroid and myeloid lineage determination, and is also required for the differentiation of lymphocytes and granulocytes during zebrafish embryogenesis.

Effects of Sunitinib Malate on Growth of Human Bladder Transitional Cell Line T24 In Vitro

Objective To investigate the growth-inhibitory effect of sunitinib malate on human bladder transitional cell carcinoma （TCC） in vitro. Methods Human bladder TCC cell line T24 was cultured and exposed to graded concentrations of sunitinib malate for 72 hours in vitro to determine the sensitivities to drug. Cell viability was measured by MTT assay. Cell apoptotic morphology was observed by fluorescence microscope following DAPl staining. Band expressions of Fas, Fas ligand, poly （ADP-ribose） polyrnerase （PARP） and D-actin were analyzed by Western blot. Wound healing process of T24 cells exposed to sunitinib malate was assayed. Results Sunitinib malate exerted a concentration-dependent and time-dependent inhibitory effect on the T24 cell lines. Fluorescence microscopy showed that small vacuoles appeared in the nuclei of T24 cells and the vacuoles were bigger with higher drug concentrations. The expressions of Fas ligand and PARP in T24 cells treated with sunitinib malate exhibited a concentration-dependent increase. Moreover sunitinib malate suppressed the wound healing process in a concentration-dependent manner. Conclusion Sunitinib malate exerted marked inhibitory activity against bladder cancer cell line T24.