Semaphorin 5A基因在胃癌侵袭和转移中的作用

目的探讨Semaphorin 5A基因在胃癌侵袭和转移中的作用机制。方法应用RNA干扰技术,构建Sema-phorin 5A-siRNA、Scrambled-siRNA表达载体,并分别转染胃癌细胞株SGC7901,建立Semaphorin 5A稳定表达抑制的胃癌细胞株SGC7901-Sema5A-si和对照组SGC7901-Scram-si胃癌细胞株;采用Western blotting、ELISA和RT-PCR方法,检测MMP2、MMP9在上述2种胃癌细胞株中的表达;应用MMP2、MMP9抗体处理胃癌细胞株,观察其对胃癌细胞株SGC7901-Sema5A-si和Semaphorin 5A侵袭、转移能力的影响。结果 SGC7901-Sema5A-si中MMP9表达水平明显低于SGC7901-Scram-si中的表达水平（P〈0.05）,MMP2在2种胃癌细胞株中表达水平无明显差异（P〉0.05）;MMP9抗体能够阻断Semaphorin 5A基因介导的胃癌侵袭和转移,MMP2抗体对Semaphorin 5A基因介导的胃癌侵袭和转移无影响。结论Semaphorin 5A可能通过MMP9表达促进胃癌发生侵袭和转移。     

Hypoxia promotes metastasis in human gastric cancer by up‐regulating the 67‐kDa laminin receptor

It has been reported that the 67‐kDa laminin receptor (67LR) is implicated in cancer metastasis. We recently showed that 37LRP, the 67LR precursor, is a hypoxia‐inducible factor 1 (HIF‐1) target gene exposed to hypoxia in gastric cancer. Here, we investigated the role of 67LR in hypoxic metastasis and invasion in gastric cancer. Immunohistochemical analysis, western blotting, and RT‐PCR assays revealed that 67LR was highly expressed in metastatic gastric cancers in vivo. Knockdown of the 67LR protein by RNA interference significantly decreased the adhesive, invasive, and in vivo metastatic abilities of the gastric cancer cell lines SGC7901 and MKN‐45. Western blot analysis showed that 67LR increased the expression of urokinase‐type plasminogen activator (uPA) and matrix metalloproteinase (MMP)‐9, and decreased tissue inhibitor of matrix metalloproteinase (TIMP)‐1 protein. We further showed that hypoxia induced 67LR expression in a time‐dependent manner and this induction was inhibited by HIF‐1 small‐interfering (si) RNA. Both ERK and JNK inhibitors significantly inhibited hypoxia‐induced expression of 67LR and the subsequent expression of uPA and MMP 9. SiRNA against 67LR or antibody against MMP9 and uPA significantly inhibited hypoxia‐induced in vitro invasive ability. Taken together, these results reveal that 67LR promotes the invasive and metastatic ability of the gastric cancer cells through increasing uPA and MMP 9 expression, with involvement of the ERK and JNK signal pathway in hypoxia‐induced 67 LR expressions and subsequent uPA and MMP9 expression. (Cancer Sci 2010)     

RhoC小干扰RNA抑制胃癌细胞的迁移和侵袭

目的：研究Ras homologyC（RhoC）在胃癌转移中的作用。方法：利用Vector NTI软件设计并构建RhoC的小干扰RNA（siRNA）载体，利用脂质体介导法将一组RhoCsiRNA分别转染胃癌SGC7901细胞。筛选出稳定抗性克隆；Westernblot检测RhoCsiRNA转染后的抑制效应，Cell Counting Kit-8检测RhoC siRNA转染细胞株的生长速度；MTT法检测转染细胞株对化疗药物的敏感性；损伤刮擦实验和TRANSWELL小室实验分别检测转染细胞株的迁移与侵袭能力。结果：在计算机辅助下成功地设计并构建了5个RhoC的小干扰RNA载体，Western blot显示其中一个RhoC的小干扰RNA RhloC-s362对RhoC的表达有明显的抑制作用；尽管下调RhoC的表达对胃癌细胞的生长和药物敏感性元明显影响，但可显著抑制肿瘤细胞的迁移与侵袭。结论：RhoC siRNARhoC-s362能够明显抑制胃癌SGC7901细胞中RhoC的表达，并进而抑制胃癌细胞的迁移与侵袭，提示RhoC可能在胃癌转移中发挥重要作用。     

CDKL1对胃癌细胞迁移及侵袭的作用

目的:探讨细胞周期素依赖激酶样蛋白1(CDKL1)对胃癌细胞系SGC7901迁移及侵袭的作用.方法:将胃癌细胞系SGC7901分为实验组及对照组,采用慢病毒转染siRNA至胃癌细胞构成实验组,转染无意义序列作为对照组.通过细胞划痕实验检测胃癌SGC7901细胞的迁移能力;Transwell小室实验检测其侵袭能力;Western blot检测CDKL1低表达后对胃癌细胞中AEG-1和MMP-9蛋白表达的影响.结果:与对照组相比,CDKL1低表达后实验组细胞迁移及侵袭能力明显降低(P＜0.05),实验组胃癌细胞中AEG-1和MMP-9蛋白的表达水平明显下降(P＜0.05).结论:特异性下调CDKL1基因可抑制胃癌细胞的迁移及侵袭能力,表明CDKL1基因可促进胃癌细胞的侵袭以及转移,并且与AEG-1和MMP-9密切相关.     

LncRNA-MALAT1在缺氧诱导胃癌侵袭转移中的作用

目的:探讨LncRNA-MALAT1对缺氧诱导胃癌侵袭转移的影响。方法:RT-PCR检测胃癌组织标本和胃癌细胞株中MALAT1的表达;通过基因重组方法构建MALAT1的siRNA载体,并感染人SGC7901及MKN45胃癌细胞,RT-PCR验证siRNA干扰有效;Transwell实验研究其对胃癌细胞SGC7901及MKN45侵袭转移能力的影响。结果:MALAT1在胃癌组织和胃癌细胞株中的表达高于癌旁组织和胃正常上皮细胞;缺氧能够促进SGC7901及MKN45细胞的侵袭转移能力,而下调MALAT1能够明显抑制缺氧条件下SGC7901及MKN45细胞的侵袭转移能力。结论:MALAT1在促进胃癌细胞的侵袭转移中发挥重要作用,为胃癌靶向治疗提供理论依据。     

丹参酮ⅡA对胃癌细胞迁移和侵袭能力的抑制作用及机制

背景与目的：有研究证实,丹参酮ⅡA (tanshinone ⅡA)对肿瘤细胞具有抑制增殖、诱导分化和促凋亡的作用,并可抑制骨肉瘤细胞迁移和侵袭。但丹参酮ⅡA抑制胃癌侵袭和转移的机制尚不明确。本研究主要探讨丹参酮ⅡA对人胃癌SGC7901细胞体外迁移和侵袭的影响。方法：不同浓度(0.5、1、2、4 μg/mL)丹参酮ⅡA分别作用体外培养的胃癌SGC7901细胞24、48、72 h后,MTT比色法检测细胞增殖活力的改变;细胞划痕实验观察细胞的迁移能力的改变;3D侵袭实验观察细胞侵袭能力的改变;Real-time PCR和蛋白质印迹法(Western blot)分别检测细胞间黏附分子1(ICAM-1)、基质金属蛋白酶-2(MMP-2)、基质金属蛋白酶-9(MMP-9)、金属蛋白酶抑制剂-2(TIMP-2)mRNA和蛋白的表达水平改变。结果：1、2、4 μg/mL丹参酮ⅡA对胃癌细胞株SGC7901有明显的抑制作用,且抑制作用存在时间-剂量依赖性(P<0.05);2 μg/mL丹参酮ⅡA呈时间依赖性抑制SGC7901细胞迁移;1、2、4 μg/mL丹参酮ⅡA呈浓度依赖性抑制SGC7901细胞侵袭;丹参酮ⅡA下调SGC7901细胞ICAM-1、MMP-2、MMP-9表达,同时可上调TIMP-2表达(P<0.05)。结论：丹参酮ⅡA可抑制胃癌SGC7901细胞的迁移和侵袭,上调TIMP-2的表达,下调ICAM-1、MMP-2、MMP-9的表达,可能是其作用机制之一。     

Gab2通过调节EMT促进胃癌的侵袭转移

目的 探讨Gab2在胃癌组织中的表达情况及其对胃癌细胞侵袭转移能力的影响与机制。方法 采用免疫组织化学SP法检测胃癌组织中Gab2、E-cadherin、N-cadherin及MMP-9的表达并分析其与淋巴结转移的关系。应用小RNA干扰技术,转染SGC7901细胞株,RT-PCR法检测瞬时转染后Gab2基因表达情况,体外侵袭实验检测细胞转染后侵袭能力的变化,Western blot法检测各蛋白的表达。结果 胃癌组织中Gab2的表达水平与淋巴结转移显著相关（P=0.002）,与E-cadherin的表达负相关（r=-0.693, P=0.000）,而与N-cadherin和MMP-9的表达水平正相关（r=0.407, P=0.021; r=0.335, P=0.017）。小RNA干扰技术降低Gab2蛋白的表达后SGC7901细胞侵袭转移能力降低,同时E-cadherin表达增多,N-cadherin和MMP-9蛋白表达降低。结论 Gab2可能通过调节EMT在胃癌侵袭转移中发挥重要作用。     

lncRNA-uc003uxs在缺氧诱导胃癌侵袭转移中的作用

目的:探讨长链非编码RNA（long non-coding RNA,lncRNA）-uc003uxs在缺氧诱导胃癌侵袭和转移中的作用。方法:三个胃癌细胞系SGC7901、MKN45、MKN28分别经常氧和缺氧孵育 24 h ,提取3例配对的胃癌细胞总RNA,利用高通量lncRNA芯片比较它们之间的表达谱差异,初步筛选与缺氧诱导胃癌侵袭转移相关的关键分子。RT-PCR法检测 lncRNA-uc003uxs在缺氧诱导的胃癌细胞（相对于常氧诱导的胃癌细胞）以及20对胃癌组织（相对于癌旁组织）中的表达水平。通过慢病毒转染,稳定下调SGC7901和MKN28细胞中lncRNA-uc003uxs的表达。通过Transwell迁移和侵袭实验以及裸鼠尾静脉注射内脏转移实验检测lncRNA-uc003uxs下调后对胃癌细胞侵袭和转移能力的影响。结果:高通量芯片分析结果显示:与常氧诱导的胃癌细胞相比,缺氧诱导的胃癌细胞 SGC7901、MKN45和MKN28中有84个共同上调的lncRNA分子以及70个共同下调的lncRNA分子,而多重筛选策略则提示:lncRNA-uc003uxs可能是缺氧诱导胃癌侵袭转移的关键lncRNA分子之一。RT-PCR结果表明:lncRNA-uc003uxs在缺氧诱导的胃癌细胞SGC7901、MKN45和MKN28中显著上调,其在胃癌组织中的表达水平也显著高于癌旁组织。Transwell实验结果显示:缺氧能够显著增加SGC7901和MKN28细胞的迁移和侵袭能力,而下调lncRNA-uc003uxs的表达后,两种细胞的迁移和侵袭能力明显下降。此外,裸鼠尾静脉内脏转移实验也证实lncRNA-uc003uxs的下调抑制了胃癌细胞SGC7901的体内肝肺转移能力。结论:利用高通量芯片筛选,在胃癌细胞中发现了一系列缺氧相关的lncRNA分子。临床标本分析及功能缺失试验证实:lncRNA-uc003uxs是一个缺氧诱导胃癌侵袭转移的关键lncRNA分子。     

Anti-cancer activity of DHA on gastric cancer—an in vitro and in vivo study

Treatment of gastric cancer remains a major challenge, and new anticancer drugs are urgently required. This study investigated whether dihydroartemisinin (DHA), a semi-synthetic derivative of artemisinin, could inhibit the growth of gastric cancer both in vitro and in vivo. A series of in vitro experiments including MTT, colony-forming, wound healing, invasion, cell cycle, cellular senescence, and apoptosis assays were performed to examine the antiproliferative and antimetastatic effects of DHA on three gastric cancer cell lines, SGC-7901, BGC823, and MGC803. The result showed that the proliferation rate and colony-forming abilities of gastric cancer cells were significantly suppressed by DHA together with significant suppression of the expressions of proliferation markers (PCNA, cyclin E, and cyclin D1), and upregulation of p21 and p27. Moreover, DHA induced cellular senescence, G1 phase cell cycle arrest and hindered the migration and invasion of gastric cancer cells corresponding with downregulation of MMP-9 and MMP-2. Furthermore, DHA significantly induced apoptosis through suppressing Bcl-2 as well as activating caspase-9 and PARP. Treatment of gastric cancer cells with DHA increased miR-15b and miR-16 expression, caused a downregulation of Bcl-2, resulting in apoptosis of gastric cancer cells. In vivo, our data showed that DHA significantly inhibited the growth of SGC7901 cell-transplanted tumors. In summary, we have shown that DHA is able to inhibit the growth and metastasis of human gastric cancer. The modulation of miR-15b and miR-16 mediated the apoptosis effects of DHA in gastric cancer cells. Our work suggested that DHA has significant anticancer effects against gastric cancer both in vivo and in vitro, indicating that it is a promising therapy for human gastric cancer.     

RNAi-mediated silencing of CD147 inhibits tumor cell proliferation,invasion and increases chemosensitivity to cisplatin in SGC7901 cells in vitro

Background

CD147 is a widely distributed cell surface glycoprotein that belongs to the Ig superfamily. CD147 has been implicated in numerous physiological and pathological activities. Enriched on the surface of many tumor cells, CD147 promotes tumor growth, invasion, metastasis and angiogenesis and confers resistance to some chemotherapeutic drugs. In this study, we investigated the possible role of CD147 in the progression of gastric cancer.

Methods

Short hairpin RNA (shRNA) expressing vectors targeting CD147 were constructed and transfected into human gastric cancer cells SGC7901 and CD147 expression was monitored by quantitative realtime RT-PCR and Western blot. Cell proliferation, the activities of MMP-2 and MMP-9, the invasive potential and chemosensitivity to cisplatin of SGC7901 cells were determined by MTT, gelatin zymography, Transwell invasion assay and MTT, respectively.

Results

Down-regulation of CD147 by RNAi approach led to decreased cell proliferation, MMP-2 and MMP-9 activities and invasive potential of SGC7901 cells as well as increased chemosensitivity to cisplatin.

Conclusion

CD147 involves in proliferation, invasion and chemosensitivity of human gastric cancer cell line SGC7901, indicating that CD147 may be a promising therapeutic target for gastric cancer.     

敲低PRMT5基因表达对胃癌细胞增殖、侵袭和凋亡的影响及其机制

目的 观察蛋白质精氨酸甲基转移酶5（PRMT5）在胃癌细胞中的表达,探讨敲低PRMT5表达水平后对胃癌细胞生物学行为的影响。方法 Western blot检测胃癌细胞系MGC803、SGC7901、MKN45和人胃黏膜上皮细胞GES-1中PRMT5蛋白表达水平,转染siRNA1和siRNA2质粒敲低PRMT5的表达水平,细胞增殖和凋亡实验、Transwell实验分别检测细胞增殖能力、凋亡率和细胞的侵袭能力,Western blot检测β-catenin、Cyclin D1和Bax蛋白表达水平。结果 与GES-1细胞比较,MGC803、SGC7901和MKN45细胞中PRMT5蛋白表达水平增加（P<0.001）。敲低PRMT5表达后细胞的增殖和侵袭能力减弱,凋亡率增加（P<0.05）,β-catenin和Cyclin D1蛋白的表达降低,Bax蛋白的表达增加（P<0.001）。结论 PRMT5在胃癌细胞中表达水平升高,敲低其表达水平后可通过减弱Wnt/β-catenin信号通路转导抑制细胞的增殖和侵袭并促进细胞凋亡。     

胃癌转移相关miRNA的差异表达谱及miR-218的生物学分析

目的 探讨胃癌转移相关微小RNA（miRNA）的差异表达情况并进行miR-218的生物学分析。方法 采用实时荧光定量PCR（qPCR）及miRNA芯片法检测低转移潜能的胃癌细胞亚系（SGC7901-NM、MKN28-NM）与高转移潜能的胃癌细胞亚系（SGC7901-M、MKN28-M）间miRNA的差异表达。提取不同转移潜能胃癌细胞系和10例胃癌冰冻组织及相应的转移淋巴结中的总RNA,利用qPCR检测miR-218在不同细胞及组织中的表达情况。结果 对不同转移潜能的胃癌细胞亚系进行芯片检测发现,与SGC7901-NM细胞比较,SGC7901-M 细胞有47个分子表达下调,15个分子表达上调。与MKN28-NM细胞比较,MKN28-M细胞有41个分子表达下调,83个分子表达上调。在SGC7901-M及MKN28-M细胞中,34个分子表达均出现下降,11个分子表达均出现上升。对不同转移潜能的胃癌细胞亚系以及人永生化正常胃黏膜细胞系GES进行检测可以发现,4种不同转移潜能的胃癌细胞亚系中miR-218的表达均低于正常胃黏膜细胞系GES,差异有统计学意义（P＜0.05）,且在高转移潜能胃癌细胞亚系中miR-218的表达均低于低转移潜能胃癌细胞系,差异有统计学意义（P＜0.05）。胃癌转移淋巴结中miR-218的表达水平为0.23±0.02,低于胃癌原发灶的1.09±0.05,差异有统计学意义（P＜0.05）。结论 胃癌转移相关miRNA会出现差异表达情况,高转移潜能胃癌细胞中的miR-218表达水平上调可能与胃癌转移存在一定关系。     

miR-486-5p对胃癌细胞株SGC7901生物学行为的影响

背景与目的：既往研究表明微小RNA-486-5p(miR-486-5p)在多种肿瘤的进展中起重要作用,但其在胃癌中作用的研究较少,本研究旨在探讨miR-486-5p对胃癌细胞株SGC7901增殖、凋亡及迁移能力的影响。方法：使用实时定量PCR(quantification real-time PCR,qRT-PCR)检测胃癌细胞株SGC7901及胃黏膜上皮细胞GES-1中miR-486-5p的表达,构建miR-486-5p过表达质粒,使用脂质体法瞬时转染胃癌细胞株SGC7901,qRT-PCR检测转染细胞后miR-486-5p的表达丰度,噻唑蓝(MTT)法及流式细胞仪检测细胞的增殖及凋亡情况,Transwell小室迁移实验检测细胞的迁移能力。结果：miR-486-5p在SGC7901细胞中表达明显下调,SGC7901细胞转染miR-486-5p过表达质粒后,miR-486-5p表达明显上调,细胞增殖、迁移能力降低,凋亡率增高,差异均有统计学意义(P<0.05)。结论：miR-486-5p可抑制胃癌细胞株SGC7901的增殖和迁移。     

沉默MTA1表达对胃癌SGC7901细胞增殖和侵袭能力的影响

目的:观察转移相关基因1（MTA1）基因沉默对人胃癌细胞SGC7901增殖和迁移侵袭能力的影响,探讨 MTA1基因在胃癌发生发展中的作用。方法:利用脂质体法将靶向MTA1的小分子干扰 RNA（siRNA） 转染到胃癌细胞株 SGC7901中。运用Real time PCR 和Western blot 技术检测SGC7901细胞内MTA1的 mRNA和蛋白表达水平。CCK-8法检测细胞增殖能力,Transwell小室实验观察细胞迁移和侵袭能力。结果:MTA1 siRNA 可有效抑制SGC7901细胞MTA1基因在 mRNA 和蛋白水平上的表达（P﹤0.01）。CCK-8实验表明MTA1 siRNA 转染后细胞增殖不受影响;Transwell小室实验表明细胞迁移和侵袭能力明显下降。结论:沉默MTA1表达可抑制胃癌细胞株 SGC7901迁移和侵袭,而不影响细胞增殖。MTA1在胃癌侵袭转移过程中可能发挥重要作用。     

Effect of tissue factor on invasion inhibition and apoptosis inducing effect of oxaliplatin in human gastric cancer cell

Objective: Tissue factor (TF) is expressed abnormally in certain types of tumor cells, closely related to invasionand metastasis. The aim of this study was to construct a human gastric cancer cell line SGC7901 stably-transfectedwith human TF, and observe effects on oxaliplatin-dependent inhibition of invasion and the apoptosis induction.Methods: The target gene TF was obtained from human placenta by nested PCR and introduced into the humangastric cell line SGC7901 through transfection mediated by lipofectamine. Stably-transfected cells were screenedusing G418. Examples successfully transfected with TF-pcDNA3 recombinant (experimental group), and emptyvector pcDNA3 (control group) were incubated with oxaliplatin. Transwell chambers were used to show changein invasive ability. Caspase-3 activity was detected using a colorimetric method and annexin-V/PI doublestainingwas applied to detect apoptosis. Results: We generated the human gastric cancer cell line SGC7901/TFsuccessfully, expressing TF stably and efficiently. Compared with the control group, invasion increased, whereascaspase-3 activity and apoptosis rate were decreased in the experimental group. Conclusion: TF can enhancethe invasive capacity of gastric cancer cells in vitro. Its increased expression may reduce invasion inhibition andapoptosis-inducing effects of oxaliplatin and therefore may warrant targeting for improved chemotherapy.     

Tanshinone IIA Reverses the Malignant Phenotype of SGC7901 Gastric Cancer Cells

Backgrounds: Tanshinone IIA (TIIA), a phenanthrenequinone derivative extracted from Salvia miltiorrhizaBUNGE, has been reported to be a natural anti-cancer agent in a variety of tumor cells. However, the effect ofTIIA on gastric cancer cells remains unknown. In the present study, we investigated the influence of TIIA on themalignant phenotype of SGC7901 gastric cancer cells. Methods: Cells cultured in vitro were treated with TIIA (0,1, 5, 10 μg/ml) and after incubation for different periods, cell proliferation was measured by MTT method andcell apoptosis and cell cycling were assessed by flow cytometry (FCM). The sensitivity of SGC7901 gastric cancercells to anticancer chemotherapy was investigated with the MTT method, while cell migration and invasion wereexamined by wound-healing and transwell assays, respectively. Results: TIIA (1, 5, 10 μg/ml) exerted powerfulinhibitory effects on cell proliferation (P < 0.05, and P < 0.01), and this effect was time- and dose-dependent.FCM results showed that TIIA induced apoptosis of SGC7901 cells, reduced the number of cells in S phase andincreased those in G0/G1 phase. TIIA also significantly increased the sensitivity of SGC7901 gastric cancer cellsto ADR and Fu. Moreover, wound-healing and transwell assays showed that TIIA markedly decreased migratoryand invasive abilities of SGC7901 cells. Conclusions: TIIA can reverse the malignant phenotype of SGC7901gastric cancer cells, indicating that it may be a promising therapeutic agent.     

EIF4E基因下调对胃癌细胞的侵袭转移能力的影响及机制研究

目的:探讨特异性下调真核起始因子EIF4E对胃癌细胞侵袭转移能力的影响及可能的作用机制。方法:采用siRNA技术下调EIF4E的表达,Western blot检测转染前后EIF4E和MMP9蛋白质表达,通过Traswell小室实验检测转染前后侵袭迁移能力的差异。结果:Western blot证实,与阴性对照相比,转染EIF4E siRNA组细胞EIF4E表达明显下调,同时与阴性对照相比,转染EIF4E siRNA组细胞MMP9蛋白表达同样明显下调,Traswell小室实验证实,下调EIF4E后,MKN28细胞的体外侵袭迁移能力显著降低。结论:在胃癌MKN28细胞中存在EIF4E基因对MMP9的调控作用,EIF4E基因可能通过调控MMP9的表达参与胃癌的侵袭迁移行为,EIF4E有望成为肿瘤基因治疗的新靶点。     

miR-9-5p靶向FGF9抑制胃癌细胞迁移侵袭的作用机制

目的:探讨miR-9-5p在胃癌组织中的表达及对胃癌细胞株SGC7901、AGS细胞迁移和侵袭影响的作用机制。方法:real-time PCR和Western blot检测癌旁组织、胃癌组织及其细胞系中miR-9-5p和FGF9的表达；Transwell小室检测过表达miR-9-5p或敲除FGF9对SGC7901和AGS细胞迁移和侵袭能力的影响；TargetScan在线分析、双荧光素酶报告基因和Western blot实验验证miR-9-5p和FGF9的靶向关系；将转染后的SGC7901细胞分为对照（NC）组和实验（miR-9-5p）组接种于裸鼠右侧腋窝皮下，观察记录裸鼠成瘤情况并绘制生长曲线。结果:与癌旁组织相比，胃癌组织及其细胞系中miR-9-5p表达下调、FGF9表达上调（P＜0.05）；过表达miR-9-5p或敲除FGF9能抑制SGC7901、AGS细胞迁移和侵袭；TargetScan在线分析、双荧光素酶报告基因和Western blot实验表明miR-9-5p能调控FGF9表达；miR-9-5p组细胞成瘤速度较NC组慢（P＜0.05），肿瘤体积及重量小（P＜0.05）。结论:miR-9-5p在胃癌组织中表达下调，过表达miR-9-5p可抑制FGF9表达，从而抑制胃癌细胞的迁移和侵袭，减缓肿瘤生长。     

胃癌细胞基质金属蛋白酶-2、9的表达及对细胞迁移的影响

 目的 研究基质金属蛋白酶 2、9(matrix metalloproteinase 2、9, MMP-2、9)在胃癌细胞株HGC-27和SGC-7901的表达及对细胞迁 徙转移的影响。 方法 采用Real time PCR检测MMP-2、9 mRNA的含量;以ELISA检测培养上清液中 MMP-2、9的相对浓度;通过明胶酶谱法测定培养上清液中的MMP-2、9比活。用Transwell小室迁徙实验、同质黏附和异质黏附实验比较两株细胞体外迁徙黏附能力。 结果 HGC-27中 MMP-2 和MMP-9 mRNA含量高于SGC-7901。HGC-27上清液 MMP-2、9相对浓度均高于SGC-7901,且MMP-2比活高于SGC-7901,两株细胞MMP-9比活相近。HGC-27穿膜细胞数和异质黏附细胞数高于SGC-7901,同质黏附细胞数低于SGC-7901,差异均有统计学意义（P<0.05）。 结论 迁徙和异质黏附能力较强、同质黏附能力较弱的胃癌细胞HGC-27的MMP-2、9表达较强,提示MMP-2、9高表达可能促进胃癌细胞迁徙转移。     

蛇毒cystatin基因转染抗人胃腺癌细胞体外侵袭作用的研究

目的:探讨蛇毒半胱氨酸蛋白酶抑制剂(sv-cystatin)对人胃腺癌细胞SGC7901侵袭转移的抑制作用.方法:采用人工拼接方法合成蛇毒cystatin cDNA,构建pcDNA3.1/sv-cystatin真核表达质粒,经脂质体转染将pcDNA3.1/sv-cystatin质粒和pcDNA3.1质粒分别导入胃腺癌细胞系SGC7901:利用RT-PCR和Western blot检测SGC7901细胞中sv-cystatin基因的表达;应用细胞-基质粘附实验、细胞运动实验及重建基底膜侵袭实验分析svcystatin表达对SGC7901细胞粘附、运动和侵袭能力的影响.结果:转染sv-cystatin基因后,SGC/sv-cystatin细胞克隆中可检测到sv-cystatin的明显表达,SGC/sv-cystatin细胞的运动能力和体外穿越重建基底膜的能力明显低于转染空载体细胞和未转染的SGC7901细胞,但其体外粘附能力未见明显变化.结论:sv-cystatin基因的表达可使胃癌SGC7901细胞体外运动能力及侵袭能力明显减弱,提示sv-cystatin具有抑制胃癌细胞侵袭转移的作用.