沉默VEGF基因对腺样囊性癌细胞侵袭的影响

目的:利用RNAi技术沉默腺样囊性癌细胞株ACC-2中VEGF的表达,探讨其对肿瘤细胞侵袭能力的影响。方法:利用RNAi技术沉默VEGF表达,用Transwell侵袭实验检测细胞体外侵袭能力的变化。结果:与对照组比较,VEGF的siRNA能有效地抑制实验组ACC-2细胞的侵袭能力。结论:沉默VEGF减弱腺样囊性癌的侵袭能力。     

CDH12对涎腺腺样囊性癌细胞侵袭能力的影响

 目的探讨CDH12基因对涎腺腺样囊性癌细胞（SACC）侵袭能力的影响。方法应用巢式RT-PCR法从涎腺腺样囊性癌高转移细胞株SACC-M的总RNA中扩增出CDH12cDNA;应用AdEasyTMXL 腺病毒载体系统构建含人CDH12基因的重组腺病毒表达载体;转染SACC-M细胞后,采用Western blot 检测CDH12蛋白分子的表达,采用侵袭小室法检测CDH12对SACC-M细胞侵袭能力的影响。结果CDH12基因重组腺病毒及空腺病毒经鉴定正确;Western blot检测发现, CDH12基因重组腺病毒转染细胞（Ad-CDH12/SACC-M）组CDH12的表达水平明显高于空腺病毒载体转染（Ad0/SACC-M）对照组;与Ad0/SACC-M 组相比,AdCDH12/SACC-M组细胞的侵袭能力明显高于Ad0/SACC-M组（ P<0.01）。结论CDH12能够促进涎腺腺样囊性癌细胞的体外侵袭能力,提示该基因在涎腺腺样囊性癌的侵袭转移中可能具有重要意义。     

阻断黏着斑激酶表达对涎腺腺样囊性癌细胞侵袭的影响

目的:研究阻断黏着斑激酶(FAK)的表达对涎腺腺样囊性癌肺高转移性细胞系(SACC-LM)侵袭的影响,并探讨其相关机制.方法:选取处于对数生长期的SACC-LM细胞随机分为空白对照组、DMSO溶媒组和Genistein组,应用Transwell小室通过平均穿膜细胞数观察Genistein 对SACC-LM细胞侵袭的影响...     

Ebp1过表达抑制腺样囊性癌细胞的体外迁移能力

目的 探讨Ebp1对人腺样囊性癌(adenoid cystic carcinoma,ACC)细胞迁移的影响及其分子机制.方法 构建Ebp1表达载体pcDNA3.1-Ebp1,建立Ebp1稳定表达细胞系ACC-Ebp1作为Ebp1组,同时设对照组和空载体组.采用细胞体外运动实验系统观察Ebp1对ACC细胞体外运动能力的影...     

CDH12基因siRNA对涎腺腺样囊性癌细胞侵袭力的影响

目的:探讨钙粘素12(Cadherin 12,CDH12)对涎腺腺样囊性癌(Salivary adenoid cystic carcinoma,SACC)细胞侵袭和转移能力的影响.方法:以人涎腺腺样囊性癌高转移细胞株(Salivary adenoid cystic carcinoma cell line with hi...     

熊果酸人腺样囊性癌对ACC-83侵袭与转移的影响

目的探讨熊果酸对人腺样囊性癌ACC-83细胞体外侵袭及转移的影响。方法应用不同浓度熊果酸处理人腺样囊性癌ACC-83细胞,通过Transwell小室模型测定其侵袭力及粘附力,细胞迁移实验测定细胞的运动能力,光学显微镜下观察细胞形态学变化。结果熊果酸能有效抑制人腺样囊性癌ACC-83细胞侵袭力、粘附力及运动能力（P〈0.05）,呈剂量效应关系（P〈0.05）。同时发现熊果酸处理组悬浮细胞增多,细胞体积变小,形态较圆,伪足数目较少。结论熊果酸具有抑制人腺样囊性癌ACC-83细胞侵袭与转移的作用。     

survivin基因沉默对腺样囊性癌细胞生长和凋亡的影响

目的 探讨RNA干扰下调survivin基因表达后腺样囊性癌细胞生物学行为变化及其与caspase-3基因表达的相关性.方法 设计、合成小干扰RNA(small interfering RNA,siRNA),构建表达载体pGenesil-shRNA-survivin,将重组质粒导入人腺样囊性癌ACC-2细胞株,G418筛选阳性克隆.采用逆转录-聚合酶链反应(RT-PCR)、Western blot分别检测转染后的ACC-2细胞中survivin、caspase-3 mRNA及蛋白表达的变化,用噻唑蓝(MTT)法检测细胞的增殖,原位细胞凋亡(TUNEL)法及透射电子显微镜研究细胞凋亡.结果 测序证实表达载体构建成功,在pGenesil-shRNA-survivin的ACC-2细胞株中,survivin mRNA及蛋白表达明显降低(P＜0.05),caspase-3 mRNA及蛋白表达明显增加(P＜0.05),survivin与easpase-3的mRNA和蛋白表达呈显著负相关(r=-0.333,P＜0.05).ACC-2细胞增殖受到抑制,透射电镜及TUNEL法可见细胞出现凋亡.结论 成功构建了survivin基因的RNAi真核表达载体,pGenesil-shRNA-survivin重组质粒能有效抑制腺样囊性痛细胞生长.     

腺样囊性癌神经侵袭动物模型建立的可行性和特异性研究

目的:建立腺样囊性癌(ACC)神经侵袭的动物模型.方法:将5×105人涎腺腺样囊性癌ACC-M细胞注射入30只裸鼠一侧眶下区皮下,分别于7、11、14、21、28 d、动物自行死亡时或56 d采集标本,H-E染色观察肿瘤能否侵袭眶下神经并测量神经侵袭的长度.另取16只裸鼠,随机分为2组,分别将ACC-M和人舌癌Tca-8113细胞注射入裸鼠眶下点皮下,14 d后H-E染色观察肿瘤生长和眶下管内神经侵袭的情况,比较两组神经侵袭的发生率.结果:30只裸鼠模型中23只出现了神经侵袭,7、11、14、21、28 d和自行死亡组神经侵袭长度分别为0.09、(0.27±0.02)、(0.38±0.05)、(0.68±0.18)、(1.07±0.36)和(2.10±0.77) mm.另16只裸鼠中8只ACC-M细胞注射裸鼠神经侵袭发生率为100%,远高于Tca-8113细胞注射裸鼠的12.5%(P<0.01).结论:该模型具备良好的可行性、特异性,是腺样囊性癌神经侵袭的良好研究平台.     

纤维连接蛋白基因EDA片段在腺样囊性癌细胞中的表达

目的通过纤维连接蛋白(FN)基因EDA片段在不同转移潜能腺样囊性癌(SACC)细胞中的表达,研究EDA片段与SACC细胞生物学特性的关系。方法应用免疫组织化学及半定量反转录聚合酶链反应(RT-PCR)法,分别从蛋白水平及mRNA水平对高、低转移SACC细胞株中FN基因EDA片断进行检测。结果高转移SACC细胞株(SACC-LM)中FN基因EDA片段蛋白及mRNA的表达高于低转移细胞株(SACC-83)。结论FN基因EDA片段的表达程度与SACC细胞不同转移潜能密切相关。     

正丁酸钠对涎腺腺样囊性癌细胞株ACC-2侵袭、迁移的影响及机制研究

目的：探讨不同浓度正丁酸钠对涎腺腺样囊性癌细胞株ACC‐2侵袭、迁移的影响与其作用机制。方法噻唑蓝（M T T ）法探索正丁酸钠作用ACC‐2细胞的最佳浓度,Transwell小室实验检测正丁酸钠对 ACC‐2细胞侵袭、迁移能力的影响, Western blot和实时荧光定量 PCR（RT‐PCR）分别检测5组浓度药物作用后ACC‐2细胞中高迁移率族蛋白1（HMGB1）、TLR4的mRNA和蛋白的表达。结果与对照组相比,0．625、1．250、2．500、5．000、10．000 mmol／L 5组浓度的正丁酸钠均能抑制ACC‐2细胞增殖（P＜0．05）且呈明显浓度依赖性;5组浓度正丁酸钠对细胞侵袭能力的影响差异无统计学意义（P＞0．05）;2．500、5．000、10．000 mmol／L浓度正丁酸钠可以抑制ACC‐2细胞的迁移能力（P＜0．05）,同时降低ACC‐2细胞 HMGB1、TLR4的mRNA及蛋白表达（P＜0．05）;相关性分析显示TLR4蛋白表达的降低与 HMGB1的抑制呈正相关（r＝0．810,P＜0．05）。结论正丁酸钠能够抑制ACC‐2细胞增殖,浓度较高时可以抑制ACC‐2细胞的迁移能力,同时降低HMGB1、TLR4的mRNA与蛋白表达,提示NaB对ACC‐2迁移能力抑制可能通过下调 HMGB1、TLR4的mRNA及蛋白表达来实现。     

RNAi knockdown of C-erbB2 expression inhibits salivary gland adenoid cystic carcinoma SACC-83 cell growth in vitro

Objective: To knockdown the C-erbB2 gene in salivary gland adenoid cystic carcinoma SACC-83 cells using RNA interference, and determine the effect of silencing C-erbB2 on cell proliferation. Methods: C-erbB2-siRNA was transfected into SACC-83 cells. RT-PCR and immunohistochemistry were used to detect C-erbB2 expression in SACC-83 cells. Cell proliferation was measured by the MTT assay and gene knockdown was achieved by RNA interference. Apoptosis was analyzed by flow cytometry. Results: Compared with the control, C-erbB2 mRNA expression was decreased in the C-erbB2-siRNA transfection group, and immunohistochemical analysis indicated that C-erbB2 protein expression was decreased. After C-erbB2-siRNA was transfected for 48 h, absorbance at 570 nm (MTT) was 0.185±0.021 compared with 0.354±0.034, 0.299±0.053, and 0.314±0.049 in the blank control, liposome control and negative control siRNA groups, respectively. The differences were statistically significant (P < 0.05) between the C-erbB2-siRNA group and the control groups. Following the C-erbB2 knockdown, the percentage of apoptotic cells was 5.63% compared with 2.04%, 2.85%, and 2.98% in the three control groups, respectively. Proliferation of SACC-83 cells was inhibited, and early apoptotic cells were increased. Conclusion: RNA interference can effectively silence C-erbB2 gene expression and inhibit growth of SACC-83 cells, which indicates the potential of targeting this gene as a novel gene therapy approach for the treatment of salivary gland adenoid cystic carcinoma.     

RNAi knockdown of C-erbB2 expression inhibits salivary gland adenoid cystic carcinoma SACC-83 cell growth in vitro

Objective：To knockdown the C-erbB2 gene in salivary gland adenoid cystic carcinoma SACC-83 cells using RNA interference,and determine the effect of silencing C-erbB2 on cell proliferation.Methods：C-erbB2-siRNA was transfected into SACC-83 cells.RT-PCR and immunohistochemistry were used to detect C-erbB2 expression in SACC-83 cells.Cell proliferation was measured by the MTT assay and gene knockdown was achieved by RNA interference.Apoptosis was analyzed by flow cytometry.Results：Compared with the control,C-erbB2 mRNA expression was decreased in the C-erbB2-siRNA transfection group,and immunohistochemical analysis indicated that C-erbB2 protein expression was decreased.After C-erbB2-siRNA was transfected for 48 h,absorbance at 570 nm （MTT）was 0.185±0.021 compared with 0.354±0.034,0.299±0.053,and 0.314±0.049 in the blank control,liposome control and negative control siRNA groups,respectively.The differences were statistically significant （P〈0.05）between the C-erbB2-siRNA group and the control groups.Following the C-erbB2 knockdown,the percentage of apoptotic cells was 5.63%compared with 2.04%,2.85%,and 2.98%in the three control groups,respectively.Proliferation of SACC-83 cells was inhibited,and early apoptotic cells were increased.Conclusion： RNA interference can effectively silence C-erbB2 gene expression and inhibit growth of SACC-83 cells,which indicates the potential of targeting this gene as a novel gene therapy approach for the treatment of salivary gland adenoid cystic carcinoma.     

IRF-4结合蛋白对人涎腺腺样囊性癌细胞增殖的抑制作用

目的 探讨IRF-4结合蛋白(IRF-4 binding protein,IBP)对人涎腺腺样囊性癌细胞增殖的影响.方法 用免疫组化(S-P法)染色技术对27例人涎腺腺样囊性癌组织和20例正常涎腺组织标本进行IBP表达的检测和分析.用脂质体法转染IBP过表达及空白对照载体至人腺样囊性癌细胞株ACC2,G418筛选后建立...     

nm23在涎腺腺样囊性癌中的表达及意义

目的研究肿瘤转移抑制基因nm23在涎腺腺样囊性癌（Salivary adenoid cystic carcinoa,SACC）中的表达,并探讨与SACC的临床特点及转移的关系。方法采用免疫组化通用型二步法检测30例SACC中nm23的表达,分析nm23基因表达变化与涎腺腺样囊性癌的病理特点及其转移的关系。结果12例nm23基因阴性表达病例中11例发生转移,肿瘤转移率91.7%,而18例nm23基因阳性表达病例中5例发生转移,阳性表达者肿瘤转移率仅为38.4%,两者比较有显著性差异（P〈0.05）。经检验等出nm23基因阴性表达者与腺样囊性癌转移有显著相关性,即nm23基因阴性表达者易发生转移,而阳性表达时不易发生转移。并发现nm23基因与患者的年龄、性别、病理类型及肿瘤大小等无关。结论nm23在口腔腺样囊性癌转移中起重要的作用。     

涎腺腺样囊性癌CKs及MSA免疫组化观察

目的：应用一组抗体探讨涎腺腺样性癌（ＡＣＣ）的免疫组化特征。方法：通过ＡＢＣ免疫组化染色检测了３０例涎腺ＡＣＣ中５４Ｋｄ及６６／５７Ｋｄ细胞角蛋白（ＣＫｓ）的分布。结果：ＡＣＣ基本上由５４Ｋｄ阳性细胞和ＭＳＡ阳性细胞构成,这两类细胞不同比例的肿瘤性增生形成ＡＣＣ的不同组织学构型。结论：ＡＣＣ免疫组化表型与正常省腺组织中的导管／腺泡单位的表型特征相似。