卵巢癌中SMS1对细胞增殖、侵袭和迁移的影响

目的探讨沉默神经鞘磷脂合成酶1(sphingomyelin synthase 1, SMS1)对卵巢癌细胞HO8910增殖、侵袭和迁移的影响。方法体外培养卵巢癌HO8910细胞株,设计siRNA-SMS1序列,转染细胞,分为阴性对照组(negative control, NC)、空白对照组(blank control, BC)、siRNA组;利用qRT-PCR技术检测各组细胞SMS1 mRNA相对表达量。MTT法检测HO8910细胞增殖情况、划痕实验检测转染后HO8910细胞迁移能力。Transwell实验检测细胞侵袭能力。薄层层析法检测HO8910细胞中SMS1活性、Cer水平。酸性神经鞘磷脂(sphingomyelin, SM)酶试剂盒、卵磷脂(phosphatidylcholine, PC)试剂盒分别检测HO8910细胞中SM、PC水平。结果卵巢癌细胞中SMS1 mRMA及蛋白相对表达量比正常卵巢细胞显著升高(P0.05);转染48 h后,siRNA组细胞中SMS1 mRMA及蛋白相对表达量、SMS1酶活性比NC组与BC组均显著降低(P0.05);siRNA-SMS1组A570比BC组与NC组显著降低(P0.05);迁移指数、侵袭细胞数较BC组与NC组显著降低(P0.05);与NC、BC组相比,siRNA组HO8910细胞中SM水平显著降低(P0.05),Cer水平显著升高(P0.05)。结论 SMS1基因沉默可抑制卵巢癌HO8910细胞增殖、迁移及侵袭能力,可能与下调细胞中SM水平、上调Cer水平有关。     

SIRT2沉默对胃癌SGC-7901细胞增殖、迁移和侵袭的影响

目的探究沉默信息调节蛋白2(SIRT2)对人胃癌SGC-7901细胞增殖、迁移和侵袭的影响及机制。方法采用实时荧光定量PCR(q RT-PCR)检测SIRT2在胃癌SGC-7901细胞和正常胃上皮GES-1细胞中的表达。采用RNA干扰技术处理SGC-7901细胞,SIRT2沉默组转染靶向SIRT2的干扰小RNA(SIRT2-siRNA1组、SIRT2-siRNA2组),阴性对照组转染SIRT2-si RNA阴性对照序列,空白对照组不转染。CKK-8法和平板细胞克隆形成实验检测SGC-7901细胞增殖, Transwell法检测SGC-7901细胞侵袭迁移能力,蛋白印迹检测SGC-7901细胞中SIRT2、丝氨酸/苏氨酸蛋白激酶(Akt)和磷脂酰肌醇-3激酶(PI3K)蛋白表达。结果人胃癌细胞SNU-1、KATO III、SGC-7901中SIRT2m RNA和蛋白水平均高于正常胃上皮GES-1细胞(P0.05)。转染SIRT2-si RNA的SGC-7901细胞中SIRT2m RNA和蛋白水平均低于阴性及空白对照组(P0.05);转染SIRT2-si RNA后,SGC-7901细胞增殖能力、克隆形成能力、迁移和侵袭能力均降低(P0.05);转染SIRT2-siRNA后SGC-7901细胞中Akt、PI3K蛋白磷酸化程度显著下降(P 0.05)。结论 SIRT2在胃癌细胞中表达上调,沉默SIRT2可能通过调节PI3K/Akt通路抑制胃癌细胞增殖、迁移和侵袭,推测SIRT2可能作为潜在靶标应用于胃癌的基因治疗。     

C1q/肿瘤坏死因子相关蛋白6(CTRP6)抑制卵巢癌细胞的增殖和迁移

目的探讨C1q/肿瘤坏死因子相关蛋白6(CTRP6)对卵巢癌细胞增殖和转移的抑制作用。方法 ELISA分别检测卵巢癌患者与健康志愿者血清,SKOV3、3AO和HO8910上皮性卵巢癌细胞与IOSE80正常卵巢上皮细胞培养上清中CTRP6的水平;采用人CTRP6重组蛋白处理高转移性卵巢癌HO8910细胞,ELISA检测HO8910细胞白细胞介素8(IL-8)和血管内皮生长因子(VEGF)的分泌水平,采用CCK-8法检测细胞增殖、TranswellTM侵袭实验检测细胞侵袭能力;采用CTRP6小干扰RNA(CTRP6 siRNA)或CTRP6抗体处理HO8910细胞,采用CCK-8法检测细胞的增殖能力、细胞划痕实验检测细胞迁移能力。结果CTRP6在卵巢癌患者血清和上皮性卵巢癌细胞培养上清中水平降低;CTRP6重组蛋白处理HO8910细胞48 h后,细胞增殖和侵袭能力显著下降;利用siRNA抑制CTRP6表达后,细胞增殖和迁移能力显著增强;CTRP6抗体处理后,细胞增殖和迁移能力亦显著增强。结论 CTRP6在卵巢癌患者血清和上皮性卵巢癌细胞上清中水平降低;CTRP6可阻断IL-8/VEGF通路从而抑制上皮性卵巢癌细胞的增殖和迁移。     

MiR-610通过下调双微体基因2的表达抑制卵巢癌细胞的侵袭和迁移

目的：探讨miR-610 对卵巢癌细胞侵袭和迁移能力的作用及机制。方法：通过RT-PCR 检测卵巢癌组织及细 胞、正常组织及细胞中miR-610 与鼠双微体基因2（MDM2）mRNA的表达;通过Lipofectamine 2000 将miR-NC、 miR-610 mimic、miR-6101 inhibitor、pc-MDM2质粒分别或联合转染进入HO8910细胞;划痕实验检测细胞迁移 能力,Transwell 法检测细胞侵袭能力,免疫印迹检测E- 钙黏着蛋白（E-cadherin）、N- 钙黏着蛋白（N-cadherin） 和MDM2蛋白表达水平,双荧光素酶报告基因检测miR-610 与MDM2靶向关系。 结果：与正常卵巢组织和上皮 细胞系HOSEpiC相比,人宫颈癌组织和细胞系中miR-610 表达明显下调,选择下调效果较明显的HO8910细胞系 进行后续实验。与Control 组相比,miR-610 mimic 组卵巢癌HO8910细胞划痕闭合率明显降低、侵袭细胞数目明 显减少、E-cadherin 明显上调、N-cadherin 表达明显下调,miR-610 inhibitor 组卵巢癌HO8910细胞划痕闭合率明显 升高、侵袭细胞数目明显增多、E-cadherin 明显下调、N-cadherin 表达明显上调。MiR-610 与MDM2 3'UTR 区存在 结合位点,且miR-610 靶向下调MDM2 表达。共转染pc-MDM2逆转了miR-610 mimic 对卵巢癌HO8910细胞迁移 和侵袭能力抑制作用。 结论：MiR-610 通过靶向下调MDM2抑制卵巢癌HO8910细胞迁移和侵袭能力。     

LAP3调控MMP-2/9的表达对人皮肤恶性黑色素瘤细胞侵袭转移的影响

目的 探讨亮氨酸氨肽酶3(LAP3)基因沉默对人皮肤恶性黑色素瘤细胞增殖、侵袭及其迁移的影响.方法 在黑色素瘤细胞中转染LAP3 siRNA,实验分为空白对照组、阴性对照组、LAP3干扰组,采用克隆形成实验、Transwell小室和划痕实验分别检测黑色素瘤细胞的增殖、侵袭和迁移能力,免疫印迹法检测黑色素瘤细胞中LAP3、MMP-2/9、上皮型钙黏蛋白、N-钙黏附素、波形蛋白的表达.结果 在LAP3干扰后,黑色素瘤细胞A2058的存活率、克隆形成率、迁移细胞数、侵袭细胞数以及LAP3、MMP-2/9、N-钙黏附素、波形蛋白的表达水平均显著降低(P<0.05),而细胞中上皮型钙黏蛋白的表达水平显著升高(P<0.05).结论 LAP3基因沉默可抑制黑色素瘤细胞的增殖,并通过减弱下游蛋白MMP-2/9的表达以及细胞上皮间质转化进程协同抑制黑色素瘤细胞侵袭和迁移.     

藏红花水提取物通过调控ERK、Bcl-2、Bax蛋白表达影响人卵巢癌细胞HO-8910增殖北大核心CSCD

目的:探讨藏红花水提取物调控ERK、Bcl-2、Bax蛋白表达对人卵巢癌细胞HO-8910增殖、凋亡、侵袭的影响。方法:以人卵巢癌细胞HO-8910为研究对象,给予不同浓度(0、0.4、0.8、1.0 mg/L)藏红花水提取物进行培养,MTT法检测细胞HO-8910的增殖活力;流式细胞术观察细胞HO-8910的凋亡率;划痕法检测细胞HO-8910的迁移情况;Transwell法检测细胞HO-8910的侵袭情况;Western blot法检测细胞HO-8910的ERK、Bcl-2、Bax蛋白表达。结果:人卵巢癌细胞HO-8910的增殖活力、迁移能力及侵袭能力受藏红花水提取物的影响,随藏红花水提取物剂量增加,人卵巢癌细胞HO-8910的增殖活力、迁移能力及侵袭能力减弱;而人卵巢癌细胞HO-8910的凋亡率则相反,随藏红花水提取物剂量增加,人卵巢癌细胞HO-8910的凋亡率逐渐增加;受藏红花水提取物影响,人卵巢癌细胞HO-8910中ERK、Bcl-2蛋白的表达降低,Bax蛋白的表达增加,且藏红花水提取物剂量越高,ERK、Bcl-2蛋白表达越低,Bax蛋白表达越高。结论:藏红花水提取物能够降低卵巢癌细胞的增殖活力、迁移能力及侵袭能力,并诱导卵巢癌细胞凋亡,这可能是通过调控ERK、Bcl-2、Bax蛋白表达来实现的。     

特异siRNA下调卵巢癌H08910细胞NAC-1基因表达并抑制生长

 目的：研究小RNA干扰NAC-1 (Nucleus accumbens-1,Nac1 or NAC-1)基因表达对卵巢癌HO8910细胞增殖的影响。方法：设计、合成针对NAC-1基因的特异性小干扰RNA（small interfering RNA,siRNA）及阴性siRNA,并同时设立空白对照,然后用脂质体转染HO8910细胞。通过实时荧光定量RT-PCR 和Western印迹法分别检测转染前后HO8910细胞中NAC-1 mRNA和蛋白表达水平,MTT法和克隆形成实验检测细胞增殖率,FCM法检测细胞周期分布。结果：针对NAC-1基因的特异性siRNA均能抑制NAC-1 mRNA和蛋白的表达,其中NAC-1-siRNA-1的沉默效率最高。转染NAC-1-siRNA-1 48h后,HO8910细胞中NAC-1 mRNA和蛋白表达水平分别下调74%和81%,而且细胞增殖明显受到抑制,细胞周期被阻滞于G1期,与转染阴性siRNA 及未转染组比较,有显著性差异(P?0.05)。结论：NAC-1特异性siRNA能够有效沉默NAC-1基因表达,并显著抑制卵巢癌HO8910细胞增殖。     

沉默S100A12对甲状腺乳头状癌K1细胞增殖、侵袭与迁移的影响

目的 评价沉默S100A12基因对甲状腺乳头状癌细胞K1的增殖、侵袭与迁移能力的影响.方法 用shRNA质粒转染K1细胞沉默S100A12基因,将细胞分为shRNA-NC组和shS100A12组,通过MTT实验评价细胞增殖生长趋势,平板克隆形成实验检测细胞克隆形成数量,Transwell实验检测细胞侵袭迁移能力,细胞划痕实验评价细胞的迁移能力,Western blot实验检测K1细胞E-cadherin,N-cadherin蛋白的表达情况.结果 与shRNA-NC组比较,shS100A12组K1细胞生长趋势明显受到抑制(P＜0.01),细胞克隆形成数量明显下降(P＜0.01),通过通透膜的细胞数量明显减少(P＜0.05),细胞的迁移速率明显减慢(P＜0.01).与shRNA-NC组比较,shS100A12组细胞E-cadherin的表达增高、N-cadherin的表达下降.结论 沉默甲状腺乳头状癌K1细胞中S100A12可以降低细胞增殖、侵袭与迁移能力,S100A12可能通过诱导EMT增强甲状腺乳头状癌细胞的侵袭与迁移能力.     

Sirtuin 6对肝癌细胞增殖的影响

目的:探究sirtuin 6(SIRT6)对肝癌细胞增殖的影响。方法:RT-q PCR检测200例肝癌病人及50例健康人外周血中的SIRT6的mRNA表达水平,并将肝癌病人外周血SIRT6的mRNA表达水平与多个临床病理参数相结合进行统计学分析。Western blotting检测SIRT6在原代肝细胞、永生化肝细胞及肝癌细胞系中的蛋白表达水平。在肝癌细胞中利用shRNA干扰SIRT6基因的表达,并通过Western blotting验证沉默效率;MTS实验检测SIRT6基因沉默对肝癌细胞活力的影响。Ed U标记实验检测SIRT6基因沉默对肝癌细胞DNA合成的影响;平板集落实验检测SIRT6基因沉默对肝癌细胞集落形成能力的影响;软琼脂集落形成实验检测SIRT6基因沉默对肝癌细胞锚定非依赖生长能力的影响。结果:SIRT6在肝癌病人的外周血中表达明显增高,且SIRT6的表达量增高与肿瘤的大小、肿瘤的分级以及肿瘤的血管侵袭相关。进一步验证发现SIRT6在肝癌细胞系中表达水平较原代肝细胞PHH及永生化肝细胞MIHA增高。在2个肝癌细胞系中沉默SIRT6可以抑制肝癌细胞的活力及DNA合成能力,也可抑制肝癌细胞集落形成能力及锚定非依赖生长能力。结论:SIRT6促进肝癌细胞的增殖及恶性转化。     

三叶因子3在甲状腺乳头状癌TPC-1细胞上皮间质转化中的作用及机制

目的 探讨三叶因子3（TFF3）基因沉默对人甲状腺乳头状癌TPC-1细胞迁移、侵袭、克隆形成能力以及对上皮间质转化（EMT）的影响及机制。 方法 免疫组织化学技术检测甲状腺乳头状癌组织芯片中Snail与TFF3的表达。划痕实验、侵袭实验和克隆形成实验分别检测TFF3基因对TPC-1细胞迁移、侵袭和克隆能力的影响;实时定量聚合酶链反应（Real-time PCR）、免疫印迹法和免疫细胞化学染色检测上皮间质转化标志物及丝裂原活化蛋白激酶（MAPK）通路相关蛋白的变化。 结果 31例甲状腺乳头状癌组织中,Snail蛋白于癌细胞胞质表达,31例癌旁组织阴性表达;Snail蛋白与TFF3表达呈正相关(r=0.8450,P<0.05)。TFF3基因沉默后,人TPC-1细胞的细胞迁移、侵袭和克隆形成能力均明显降低(P<0.01);TPC-1细胞上皮间质转化标志物及MAPK通路相关蛋白细胞外调节蛋白激酶（ERK）1/2和p-ERK1/2也显著降低(P<0.05)。 结论 沉默TFF3可能通过MAPK通路相关蛋白ERK1/2抑制上皮间质转化,降低TPC-1细胞迁移、侵袭和增殖能力。     

Role of ABCG2 gene in proliferation and invasion of colorectal cancer stem cells北大核心

BACKGROUND: ABCG2 transporter can mediate multidrug resistance. ABCG2 overexpression can enhance the tolerance of stem cells to chemotherapeutic drugs. However, the mechanism of ABCG2 gene in the proliferation and invasion of colorectal cancer stem cells has not been confirmed. OBJECTIVE: To investigate the role of ABCG2 gene in the proliferation and invasion of colorectal cancer stem cells. METHODS: Colorectal cancer cell lines HCT116 were routinely isolated and cultured, and were randomly divided into four groups. CD133 positive cells were idted by immunomagnetic beads method, and were transfected with ABCG2-siRNA and ABCG2 overexpression plasmids to construct colorectal cancer stem cell modell with ABCG2 low expression and overexpression, and were set as low expression group and ovvrexpression groups, respectively. The remaining HCT116 cells were set as normal and smpty plasmid transfected control groups. The proliferation and invasion of colorectal cancer stem cell in different modell were determined by MTT assay and Transwell assay, respectively. Expressions of matrix metalloproteinase 9 (IMMP-9) mRNA snd protein were detected by enzyme linked immunosorbent sssay (ELISA) and polymerase chain reaction (PCR), respectively. RESULTS AND CONCLUSION: (1) Flow cytometry results showed that: CD133 positive cells in colorectal cancer cell line HCT116 accounted for 2.4%, while increased to 94.51 % of the cell line after being sorted by immunomagnetic beads, suggesting that isolated and cultured cell were colorectal cancer stem cell. (2) MTT assy showed no significant difference between the four groups of cell prior to the cell transfection (P > 0.05). However, MTT value in the low expression group was significantly lower than that in the overexpression group after cell transfection (P < 0.05). (3) Results from the Transwell assay showed that the cells in the low expression groups did not have the ability of migration; on the contrary, in the overexpression group, the number of cells crossing the basement membrane was relatively increased and the ability of cell migration and invasion was enhanced significantly (P <0.05). (4) The IVIMP-9 protein level was lower in the low expression group than in the overexpression group (P < 0.05). Similar results were yielded in the PCR detection. To conclude, down-regulation of ABCG2 protein expression can inhibit colorectal cancer stem cell proliferation and invasion. ABCG2 gene can change the viability of colorectal cancer stem cell by regulating the IMMP-9 expression. © 2018, Journal of Clinical Rehabilitative Tissue Engineering Research. All rights reserved.     

长链非编码RNA PVT1调控miR-551通过Wnt信号通路对卵巢癌迁移和侵袭的影响

目的:探讨长链非编码RNA PVT1在卵巢癌组织中的表达情况及其在卵巢癌细胞迁移和侵袭过程中的作用及机制。方法:q PCR检测卵巢癌和正常卵巢组织及不同卵巢癌细胞中PVT1的表达情况;Transwell侵袭实验和细胞划痕实验分别检测沉默PVT1后卵巢癌细胞侵袭和迁移能力的变化;双萤光素酶报告基因检测PVT1与微小RNA(miR)-551的相互作用;Transwell侵袭实验和细胞划痕实验分别检测沉默PVT1后miR-551-inhibitor对卵巢癌细胞侵袭和迁移能力的影响;Western blot法检测沉默PVT1后Wnt信号通路相关蛋白的表达情况。裸鼠皮下成瘤实验检测沉默PVT1对卵巢癌成瘤重量及体积的影响。结果:与正常卵巢组织相比,卵巢瘤组织中PVT1表达明显增高(P0.05);卵巢癌细胞株ES-2中PVT1表达水平最高(P0.05);沉默PVT1可以抑制卵巢癌细胞侵袭和迁移能力;PVT1能与miR-551的位点特异性结合;沉默PVT1后,miR-551-inhibitor可以促进卵巢癌细胞侵袭和迁移能力;沉默PVT1后Wnt信号通路蛋白的表达相应下调;与阴性对照组相比,PVT1-siRNA组荷瘤小鼠肿瘤体积和重量都明显减小(P0.05)。结论:PVT1在卵巢癌发生发展过程中起重要作用,它可以靶向调节miR-551,通过Wnt信号通路调控卵巢癌细胞的侵袭和迁移能力。     

miRNA-22在卵巢癌组织的表达及其对卵巢癌细胞增殖、迁移与侵袭的影响

目的:观察在不同卵巢组织中miRNA-22的表达,并研究上调miRNA-22表达对卵巢癌细胞株SKOV-3增殖、迁移与侵袭的影响。方法:实时荧光定量PCR(q PCR)检测不同卵巢组织样本miRNA-22的表达;随机将SKOV-3细胞分为正常对照组(control组)、空白载体组(miRNA-22-NC组)和转染miRNA-22-mimic组(miRNA-22-m组),q PCR检测各组细胞miRNA-22的表达,CCK-8法检测细胞存活率,划痕实验与Transwell小室法分别检测细胞的迁移与侵袭能力,Western blot法检测VEGF和P53蛋白表达。结果:卵巢癌组织中miRNA-22的水平显著低于正常卵巢组织;与control组相比,miRNA-22-m组细胞miRNA-22的表达显著增加,细胞存活率显著降低,细胞迁移数和侵袭数下降,VEGF和P53的蛋白表达水平显著下降。结论:miRNA-22的低表达可能与卵巢癌的发生发展有关。过表达miRNA-22能够抑制卵巢癌细胞株的增殖、迁移和侵袭,其机制可能与下调VEGF和P53的蛋白表达有关。     

CADM1过表达对人胃癌MKN-45细胞增殖和侵袭的影响及其分子机制

 目的：研究细胞黏附分子1（cell adhesion molecule 1,CADM1）过表达对人胃癌MKN-45细胞增殖和侵袭的影响并探讨其可能的分子机制。方法：采用Western blotting检测3株胃癌细胞系中CADM1蛋白的表达。构建pcDNA-CADM1真核表达载体,并将其转染MKN-45细胞,采用G418筛选稳定表达CADM1的细胞株,利用Western blotting鉴定所筛选的稳定细胞株。采用CCK-8试剂和Boyden小室分析过表达CADM1对MKN-45细胞增殖和侵袭的影响。利用Western blotting检测细胞增殖和侵袭相关蛋白表达。结果：MKN-45细胞中CADM1蛋白的相对表达水平显著低于MKN-28和SGC-7901细胞（P<0.05）。此外,成功构建pcDNA-CADM1真核表达载体,并获得稳定过表达CADM1的MKN-45细胞株。CCK-8结果显示,与未处理组和pcDNA3.1组相比,pcDNA-CADM1组中MKN-45细胞的增殖明显受到抑制（P<0.05）。Boyden小室体外侵袭实验结果显示,与未处理组（101.53±6.89）和pcDNA3.1组（98.77±7.03）相比,pcDNA-CADM1组中MKN-45细胞穿膜的细胞数（52.35±3.89）显著减少（P<0.05）。Western blotting结果显示,与未处理组和pcDNA3.1组相比,pcDNA-CADM1组中p21蛋白表达显著上调,而MMP-2和MMP-9表达显著下调（P<0.05）。结论：CADM1过表达能明显抑制胃癌细胞的增殖和侵袭能力,因而CADM1有望成为胃癌治疗的新靶点。     

Biological significance of focal adhesion kinase in ovarian cancer: role in migration and invasion

Focal adhesion kinase (FAK) is a nonreceptor tyrosine kinase that is activated by integrin clustering. There are limited data regarding the functional role of FAK in ovarian cancer migration and invasion. In the current study, FAK expression was evaluated in ovarian cell lines (nontransformed and cancer), 12 benign ovarian samples, and in 79 invasive epithelial ovarian cancers. All three ovarian cancer cell lines overexpressed FAK compared to the nontransformed cells. The dominant-negative construct called FAK-related nonkinase (FRNK) was introduced into two ovarian cancer cell lines (SKOV3 and 222). FRNK promoted FAK dephosphorylation without changing total FAK levels in these cell lines. Furthermore, FRNK decreased the in vitro invasive ability of ovarian cancer cells by 56 to 85% and decreased migration by 52 to 68%. FRNK-transfected cells also displayed poor cell spreading. Immunohistochemical analysis revealed that the surface epithelium from all benign ovarian samples had weak FAK expression. In contrast, 68% of invasive ovarian cancers overexpressed FAK. FAK overexpression was significantly associated with high tumor stage, high tumor grade, positive lymph nodes and presence of distant metastasis (all P values <0.05). FAK overexpression was also associated with shorter overall survival (P = 0.008). Multivariate analysis revealed that FAK overexpression and residual disease >1 cm were independent predictors of poor survival. These data indicate that FAK is overexpressed in most invasive ovarian cancers and plays a functionally significant role in ovarian cancer migration and invasion. Thus, FAK may be an important therapeutic target in ovarian carcinoma.     

miR-107 靶向NID2 通过Notch 信号通路调控肺癌细胞的增殖侵袭

目的:miR-107 靶向NID2 调控Notch 信号通路影响肺癌的侵袭和增殖。方法:免疫组化检测NID2 在肺癌组织和正常肺组织中的表达;PCR 检测miR-107 在肺癌组织中的表达;双荧光素酶报告基因系统检测miR-107 对NID2 转录活性的影响;肿瘤细胞成球实验检测miR-107 的表达对肺癌细胞A549 的增殖能力的影响;Transwell 侵袭实验检测miR-107 的表达对肺癌A549 细胞的侵袭能力的影响;划痕试验检测miR-107 的表达对肺癌A549 细胞的迁移能力的影响;Western blot 检测过表达miR-107 后Notch 信号通路的蛋白表达水平。结果:和正常肺组织比较,NID2 在肺癌组织中表达较高;miR-107 在肺癌组织中表达明显降低;双荧光素酶报告基因系统检测结果显示,miR-107 可以直接调控NID2 的转录活性;过表达miR-107 后,肺癌细胞A549 的增殖、侵袭和迁移能力明显降低;过表达miR-107 后,Notch1、hes-1、presenilin1 蛋白表达下调。结论:miR-107靶向NID2 的表达,通过Notch 通路调控肺癌细胞的增殖和侵袭能力。     

布托啡诺调控PBX3基因抑制乳腺癌细胞系MCF7增殖、迁移和侵袭

目的探讨布托啡诺(butorphanol)对乳腺癌细胞增殖、迁移和侵袭的影响以及相关的分子机制。方法用MTT法检测不同浓度布托啡诺对人乳腺癌细胞系MCF7的抑制作用;用Transwell迁移及侵袭实验检测不同浓度布托啡诺对人乳腺癌MCF7细胞迁移及侵袭的影响;RT-qCR与Western blot法分别检测乳腺癌细胞系、正常乳腺上皮细胞以及布托啡诺对MCF7细胞中PBX3 mRNA及蛋白表达的影响;观察转染si-PBX3或si-con后,MCF7细胞增殖、迁移及侵袭能力的变化;PBX3过表达验证布托啡诺对乳腺癌增殖、迁移及侵袭的作用机制。Western blot检测cyclin D1和MMP-2蛋白表达。结果PBX3在乳腺癌细胞系中的表达上调,沉默PBX3表达可明显抑制MCF7细胞增殖、迁移及侵袭,同时抑制cyclin D1和MMP-2的表达;不同浓度的布托啡诺干预能显著抑制MCF7细胞增殖、迁移及侵袭且具有浓度依赖性,还可抑制PBX3、cyclin D1和MMP-2的表达;过表达PBX3可逆转布托啡诺对乳腺癌细胞增殖、迁移及侵袭的抑制作用。结论布托啡诺可通过抑制PBX3降低乳腺癌细胞增殖、迁移及侵袭能力。     

Role of MAPK7 in cell proliferation and metastasis in ovarian cancer

Aim: To investigate the effect of mitogen-activated protein kinase 7 (MAPK7) in ovarian cancer metastasis and to explore its potential mechanism. Methods: pcDNA-MAPK7 and siRNA-MAPK7 vectors were transfected into the human ovarian cell line OVCAR-3 based on gene silencing and overexpression methods. Effects of MAPK7 overexpression and silencing on OVCAR-3 cells proliferation, cell invasion, and migration were analyzed using the MTT (3-[4, 5-dimethylthiazol-2-yl]-2, 5-diphenyl tetrazolium bromide) assay, Matrigel methods, and Markered methods respectively. In addition, effect of MAPK7 expression on extracellular matrix (ECM) associated protein was detected using Western blot. Results: Compared with the controls, MAPK7 was up-regulated when cells were transfected with pcDNA-MAPK7 plasma, as well as MAPK7 was sliced when cells were transfected with siRNA-MAPK7 plasma (P<0.05). Besides, biological function analysis performed that overexpression of MAPK7 significantly increased OVCAR-3 cell proliferation, invasion, and migration (P<0.05), while these effects were inhibited by MAPK7 silencing (P<0.05). Additionally, MAPK7 overexpression increased type II collagen expression (P<0.05). However, there was no significant difference between MAPK7 expression and type I collagen expression (P>0.05). Conclusion: Our data implied the up-regulated MAPK7 might contribute to ovarian cancer metastasis through up-regulating type II collagen expression and then were involved in cell biological processes such as cell proliferation, invasion, and migration. MAPK7 may be a potential therapeutic target in the clinical treatment for ovarian cancer.