Long noncoding RNAs HAND2-AS1 ultrasound microbubbles suppress hepatocellular carcinoma progression by regulating the miR-873-5p/tissue inhibitor of matrix metalloproteinase-2 axis

BACKGROUND Increasing data indicated that long noncoding RNAs(lncRNAs)were directly or indirectly involved in the occurrence and development of tumors,including hepatocellular carcinoma(HCC).Recent studies had found that the expression of lncRNA HAND2-AS1 was downregulated in HCC tissues,but its role in HCC progression is unclear.Ultrasound targeted microbubble destruction mediated gene transfection is a new method to overexpress genes.AIM To study the role of ultrasound microbubbles(UTMBs)mediated HAND2-AS1 in the progression of HCC,in order to provide a new reference for the treatment of HCC.METHODS In vitro,we transfected HAND2-AS1 siRNA into HepG2 cells by UTMBs,and detected cell proliferation,apoptosis,invasion and epithelial-mesenchymal transition(EMT)by cell counting kit-8 assay,flow cytometry,Transwell invasion assay and Western blotting,respectively.In addition,we transfected miR-837-5p mimic into UTMBs treated cells and observed the changes of cell behavior.Next,the UTMBs treated HepG2 cells were transfected together with miR-837-5p mimic and tissue inhibitor of matrix metalloproteinase-2(TIMP2)overexpression vector,and we detected cell proliferation,apoptosis,invasion and EMT.In vivo,we established a mouse model of subcutaneous transplantation of HepG2 cells and observed the effect of HAND2-AS1 silencing on tumor formation ability.RESULTS We found that UTMBs carrying HAND2-AS1 restricted cell proliferation,invasion,and EMT,encouraged apoptosis,and HAND2-AS1 silencing eliminated the effect of UTMBs.Additionally,miR-873-5p targets the gene HAND2-AS1,which also targets the 3’UTR of TIMP2.And miR-873-5p mimic counteracted the impact of HAND2-AS1.Further,miR-873-5p mimic solely or in combination with pcDNA-TIMP2 had been transformed into HepG2 cells exposed to UTMBs.We discovered that TIMP2 reversed the effect of miR-873-5p mimic caused by the blocked signalling cascade for matrix metalloproteinase(MMP)2/MMP9.In vivo results showed that HAND2-AS1 silencing significantly inhibited tumor formation in mice.CONCLUSION LncRNA HAND2-AS1 promotes TIMP2 expression by targeting miR-873-5p to inhibit HepG2 cell growth and delay HCC progression.

生殖道感染对足月和未足月胎膜早破孕妇宫颈分泌液MMP-2、MMP-9表达及不良母婴结局的影响

目的分析生殖道感染对足月和未足月胎膜早破(PROM)孕妇基质金属蛋白酶-2(MMP-2)、基质金属蛋白酶-9(MMP-9)表达及不良母婴结局的影响。方法招募2021年6月至2023年6月就诊于湖北医药学院附属十堰市太和医院的110例PROM孕妇作为研究对象。根据生殖道感染情况和孕周将其分为未足月胎膜早破(pPROM)感染组(34例)、pPROM非感染组(26例)、足月胎膜早破(tPROM)感染组(34例)、tPROM非感染组(16例)。比较四组一般资料、宫颈分泌液MMP-2和MMP-9表达水平以及不良母婴结局发生情况,采用多因素logistic回归分析PROM孕妇发生不良母婴结局的影响因素。结果四组年龄、孕次、体质量指数、吸烟史、饮酒史比较差异无统计学意义(P>0.05)。四组宫颈分泌液MMP-2、MMP-9表达水平比较差异有统计学意义(P<0.05),其中pPROM感染组宫颈分泌液MMP-2、MMP-9表达水平高于pPROM非感染组和tPROM非感染组,tPROM感染组宫颈分泌液MMP-2、MMP-9表达水平高于tPROM非感染组,差异有统计学意义(P<0.05)。四组绒毛膜羊膜炎及不良母婴结局总发生率比较差异有统计学意义(P<0.05),其中pPROM感染组绒毛膜羊膜炎发生率高于tPROM感染组和tPROM非感染组,不良母婴结局总发生率高于pPROM非感染组和tPROM非感染组,差异有统计学意义(P<0.05)。多因素logistic回归分析结果显示,生殖道感染及较高的宫颈分泌液MMP-2、MMP-9表达水平是PROM孕妇发生不良母婴结局的独立危险因素(P<0.05)。结论生殖道感染可增加足月和未足月PROM孕妇宫颈分泌液MMP-2、MMP-9表达水平,生殖道感染及较高的宫颈分泌液MMP-2、MMP-9表达水平是PROM孕妇发生不良母婴结局的独立危险因素。

金刚藤胶囊联合桂枝茯苓胶囊治疗慢性盆腔炎的疗效及对血清GM-CSF、MMP-2水平的影响

【目的】分析金刚藤胶囊联合桂枝茯苓胶囊治疗湿热瘀结型慢性盆腔炎的疗效及对血清粒细胞-巨噬细胞集落刺激因子(GM-CSF)、基质金属蛋白酶2(MMP-2)水平的影响。【方法】将90例湿热瘀结型慢性盆腔炎患者随机分为联合组和单药组,每组各45例。单药组患者给予桂枝茯苓胶囊治疗,联合组患者在单药组的基础上联合金刚藤胶囊治疗,疗程为2周并随访1个月。观察2组患者治疗前后中医证候积分和血清GM-CSF、MMP-2水平的变化情况,比较2组患者的临床疗效、症状缓解时间、疾病复发和不良反应发生情况。【结果】(1)疗效方面,治疗2周后,联合组的总有效率为93.33%(42/45),单药组为66.67%(30/45),组间比较,联合组的疗效明显优于单药组(P<0.01)。(2)中医证候积分方面,治疗后,2组患者的下腹疼痛、腰骶疼痛、带下量多、月经量多、经行腹痛、神疲乏力等各项中医证候积分均较治疗前明显降低(P<0.05),且联合组对各项中医证候积分的降低幅度均明显优于单药组(P<0.05)。(3)症状缓解时间方面,联合组患者治疗后的白带恢复正常时间、下腹坠胀和腹痛缓解时间均较单药组明显缩短(P<0.01)。(4)血清学指标方面,治疗后,2组患者的血清GM-CSF、MMP-2水平均较治疗前明显降低(P<0.05),且联合组对血清GM-CSF、MMP-2水平的降低幅度均明显优于单药组(P<0.05或P<0.01)。(5)疾病复发和不良反应方面,联合组的复发率和不良反应发生率分别为11.11%(5/45)、13.33%(6/45),均明显低于单药组的35.56%(16/45),差异均有统计学意义(P<0.05或P<0.01)。【结论】金刚藤胶囊联合桂枝茯苓胶囊治疗,可以显著提高湿热瘀结型慢性盆腔炎的临床疗效,有效缩短症状缓解时间,降低血清GM-CSF、MMP-2水平。

安罗替尼联合PD-1抑制剂治疗晚期非小细胞肺癌患者的近期临床疗效及对血清VEGF、MMP-2水平的影响分析

目的分析安罗替尼联合程序性死亡受体1(PD-1)抑制剂治疗晚期非小细胞肺癌患者的近期临床疗效及对血清血管内皮生长因子(VEGF)、基质金属蛋白酶-2(MMP-2)水平的影响。方法选择2018年2月至2022年2月成都市第三人民医院收治的110例ⅢB~Ⅳ期非小细胞肺癌患者的临床资料,根据治疗方法不同将其分为联合治疗组(接受安罗替尼联合PD-1抑制剂治疗)和抗PD-1治疗组(接受PD-1抑制剂治疗),每组55例。两组均以21 d为一个治疗周期,每个治疗周期后停药7 d,共治疗3个周期。比较两组治疗前、治疗后1周、治疗后1个月、治疗后3个月的血清VEGF、MMP-2水平,以及疗程结束时的临床疗效。结果在接受治疗后,两组血清VEGF、MMP-2水平均呈下降趋势(P<0.05),且在治疗后1周、1个月、3个月,联合治疗组水平均较抗PD-1治疗组更低,差异有统计学意义(P<0.05)。联合治疗组的客观缓解率(ORR)、疾病控制率(DCR)均显著高于抗PD-1治疗组,差异有统计学意义(45.45%vs 23.64%,90.91%vs 76.36%;P<0.05)。两组总不良反应发生率比较差异无统计学意义(70.91%vs 78.18%;P>0.05)。结论安罗替尼联合PD-1抑制剂对晚期非小细胞肺癌患者疗效显著,能降低血清VEGF、MMP-2水平,且安全性较高。

伊马替尼联合重组人干扰素α-2b治疗加速期慢性髓性白血病患者的疗效及对血清MMP-2、SALL4 mRNA和AGP水平的影响

目的探讨伊马替尼联合重组人干扰素α-2b(IFNα-2b)治疗加速期慢性髓性白血病(CML)患者的疗效及对血清基质金属蛋白酶-2(MMP-2)、婆罗双树样基因4(SALL4)mRNA和α1-酸性糖蛋白(AGP)水平的影响。方法招募2017年1月至2021年1月陕西省人民医院收治的加速期CML住院患者90例。采用随机数字表法将其分为观察组(采用伊马替尼联合IFNα-2b治疗,45例)和对照组(采用伊马替尼治疗,45例)。比较两组临床疗效和血清MMP-2、SALL4 mRNA和AGP水平以及不良反应发生情况。结果观察组治疗后获完全血液学缓解(CHR)的人数比例较对照组更高,整体疗效更优,差异有统计学意义(P<0.05)。两组治疗前血清白细胞介素-6(IL-6)、前列腺素E 2(PGE 2)、碱性磷酸酶(ALP)、MMP-2、SALL4 mRNA、AGP水平比较差异无统计学意义(P>0.05),治疗后IL-6、PGE 2、ALP、MMP-2、SALL4 mRNA、AGP水平均较治疗前显著降低(P<0.05),且观察组治疗后水平较对照组更低,差异有统计学意义(P<0.05)。观察组血液学不良反应发生率高于对照组[22.22%(10/45)vs 6.67%(3/45);χ2=4.406,P=0.039],两组非血液学不良反应发生率比较差异无统计学意义[6.67%(3/45)vs 4.44%(2/45);χ2=0.000,P=1.000],总不良反应发生率比较差异有统计学意义[28.89%(13/45)vs 11.11%(5/45);χ2=4.444,P=0.035]。结论伊马替尼联合IFNα-2b较单用伊马替尼治疗加速期CML疗效更佳,可降低患者血清MMP-2、SALL4 mRNA和AGP水平,但可能会增加患者血液学不良反应的发生风险。

A matrix metalloproteinase-responsive hydrogel system controls angiogenic peptide release for repair of cerebral ischemia/reperfusion injury

Vascular endothelial growth factor and its mimic peptide KLTWQELYQLKYKGI(QK)are widely used as the most potent angiogenic factors for the treatment of multiple ischemic diseases.However,conventional topical drug delivery often results in a burst release of the drug,leading to transient retention(inefficacy)and undesirable diffusion(toxicity)in vivo.Therefore,a drug delivery system that responds to changes in the microenvironment of tissue regeneration and controls vascular endothelial growth factor release is crucial to improve the treatment of ischemic stroke.Matrix metalloproteinase-2(MMP-2)is gradually upregulated after cerebral ischemia.Herein,vascular endothelial growth factor mimic peptide QK was self-assembled with MMP-2-cleaved peptide PLGLAG(TIMP)and customizable peptide amphiphilic(PA)molecules to construct nanofiber hydrogel PA-TIMP-QK.PA-TIMP-QK was found to control the delivery of QK by MMP-2 upregulation after cerebral ischemia/reperfusion and had a similar biological activity with vascular endothelial growth factor in vitro.The results indicated that PA-TIMP-QK promoted neuronal survival,restored local blood circulation,reduced blood-brain barrier permeability,and restored motor function.These findings suggest that the self-assembling nanofiber hydrogel PA-TIMP-QK may provide an intelligent drug delivery system that responds to the microenvironment and promotes regeneration and repair after cerebral ischemia/reperfusion injury.

All-trans retinoic acid regulates the expression of MMP-2 and TGF-β2 via RDH5 in retinal pigment epithelium cells

·AIM: To investigate the effect of all-trans retinoic acid(ATRA) on retinol dehydrogenase 5(RDH5), matrix metalloproteinase-2(MMP-2) and transforming growth factor-β2(TGF-β2) transcription levels, and the effect of RDH5 on MMP-2 and TGF-β2 in retinal pigment epithelium(RPE) cells.·METHODS: After adult RPE cell line-19(ARPE-19 cells) intervened with gradient concentrations of ATRA(0-20 μmol/L) for 24h, flow cytometry was used to detect the proliferation and apoptosis of cells in each group, and quantitative realtime polymerase chain reaction(q RT-PCR) was used to detect RDH5, MMP-2 and TGF-β2 m RNA expression. Then, after ARPE-19 cells transfected with three different si RNA targets for 48h, the RDH5 knockdown efficiency of each group and expression of MMP-2 and TGF-β2 m RNA within them was detected by q RT-PCR. ·RESULTS: Flow cytometry results showed that ATRA could inhibit the proliferation of RPE cells and promote the apoptosis of RPE cells, and the difference of apoptosis was statistically significant when the ATRA concentration exceeded 5 μmol/L and compared with the normal control group(P=0.027 and P=0.031, respectively). q RT-PCR results showed that ATRA could significantly inhibit the expression level of RDH5 m RNA(P<0.001) and promote the expression of MMP-2 and TGF-β2 m RNA(P=0.03 and P<0.001, respectively) in a dose-dependent manner, especially when treated with 5 μmol/L ATRA. The knockdown efficiency of RDH5 si RNA varies with different targets, among which RDH5 si RNA-435 had the highest knockdown efficiency, i.e., more than 50% lower than that of the negative control group(P=0.02). When RDH5 was knocked down for 48h, the results of q RT-PCR showed that the expressions of MMP-2 and TGF-β2 m RNA were significantly up-regulated(P<0.001).·CONCLUSION: ATRA inhibits the expression of RDH5 and promotes MMP-2 and TGF-β2, and further RDH5 knockdown significantly upregulates MMP-2 and TGF-β2. These findings suggest that RDH5 may be involved in an epithelial-mesenchymal transition of RPE cells mediated by ATRA.

补阳还五汤联合巴曲酶对突发性耳聋患者中医证候、听力水平及血清ET-1、CysC、MMP-2的影响

【目的】探讨补阳还五汤联合巴曲酶治疗突发性耳聋的疗效及对患者中医证候、听力水平和血清血管内皮素1(ET-1)、胱抑素C(CysC)、基质金属蛋白酶2(MMP-2)水平的影响。【方法】将92例突发性耳聋患者随机分为对照组和观察组,每组各46例。对照组给予巴曲酶静脉滴注治疗,隔日1次;观察组在对照组的基础上联合补阳还五汤治疗,每日1剂;15 d为1个疗程,连续治疗3个疗程。观察2组患者治疗前后中医证候积分、听力水平及血清ET-1、CysC、MMP-2水平的变化情况,并评价2组患者的临床疗效和安全性。【结果】(1)治疗3个疗程后,观察组的总有效率为91.30%(42/46),对照组为71.74%(33/46),组间比较(χ2检验),观察组的疗效明显优于对照组(P<0.05)。(2)治疗后,2组患者的耳聋耳闷、头晕耳鸣、口苦咽干、舌红苔黄等各项中医证候积分均较治疗前降低(P<0.05),且观察组的降低作用均明显优于对照组(P<0.01)。(3)治疗后,2组患者的纯音听觉阈值(PTA)、听觉诱发电位(AEP)、听觉脑干反应(ABR)等听力指标均较治疗前改善(P<0.05),且观察组的改善作用均明显优于对照组(P<0.01)。(4)治疗后,2组患者的血清ET-1、CysC、MMP-2水平均较治疗前降低(P<0.05),且观察组的降低作用均明显优于对照组(P<0.01)。(5)治疗期间,观察组的不良反应发生率为13.04%(6/46),对照组为8.70%(4/46),组间比较,差异无统计学意义(P>0.05)。【结论】补阳还五汤联合巴曲酶治疗突发性耳聋疗效显著,能有效改善患者中医证候,提高患者听力水平,改善血清学指标,且具有较高的安全性。

卵巢癌患者MMP-2基因rs243866,rs243842和rs243865位点多态性与含铂化疗方案疗效及不良反应的相关性研究

目的探讨卵巢癌(ovarian cancer)患者基质金属蛋白酶(matrix metalloproteinase,MMP)-2基因rs243866,rs243842和rs243865的多态性与其含铂化疗方案的疗效和不良反应的关系。方法选取2018年12月~2021年12月在海南医学院第二附属医院经病理确诊、一线接受含铂化疗方案治疗并具有完整资料的卵巢癌患者126例。化疗前采集患者3ml外周静脉血,采用多重PCR扩增目标区域后再进行高通量测序,鉴定rs243866,rs243842和rs243865的基因型。分析其与含铂方案疗效及不良反应的相关性。结果109例患者可评估疗效,其中完全缓解(complete response,CR)/部分缓解(partial response,PR)或疾病稳定(stable disease,SD)/疾病进展(progressive disease,PD)的患者分别为78例和31例。126例患者中发生Ⅲ~Ⅳ级血液学毒性和消化道毒性的分别有33例和16例。MMP-2基因rs243866多态性与含铂化疗方案的疗效及不良反应均相关,携带G等位基因的卵巢癌患者具有更好的疗效,其CR/PR率为74.6%,显著高于携带A等位基因(25.4%)的患者(OR=3.148,95%CI:1.362~7.441)。携带A等位基因的患者Ⅲ~Ⅳ级血液学毒性的发生率为60%,显著高于携带G等位基因(22.5%)的患者(OR=5.176,95%CI:2.193~12.217)。rs243842和rs243865位点各基因型分布及等位基因分布频次与含铂方案的疗效及不良反应均无相关性。结论携带rs243866 G等位基因的晚期卵巢癌患者在接受含铂方案治疗时具有较好的疗效和较低Ⅲ/Ⅳ血液学毒性。

Suppression of matrix metalloproteinase-2 via RNA interference inhibits pancreatic carcinoma cell invasiveness and adhesion

AIM:To investigate the inhibitory effects of RNA interference (RNAi) on expression of matrix metalloproteinase-2 (MMP-2) gene and invasiveness and adhesion of human pancreatic cancer cell line,BxPC-3.METHODS:RNAi was performed using the vector (pGPU6)-based small interference RNA (siRNA) plasmid gene silence system to specifically knock down MMP-2 expression in pancreatic cancer cell line,BxPC-3. Four groups of different specific target sequence in coding region of MMP-2 and one non-specific sequence were chosen to construct four experimental siRNA plasmids of pGPU6-1,pGPU6-2,pGPU6-3 and pGPU6-4,and one negative control siRNA plasmid of pGPU6 (-). MMP-2 expression was measured by reverse transcription polymerase chain reaction (RT-PCR) and Western blot. Cell proliferation and apoptosis were examined by methyl thiazolyl tetrazolium (MTT) and flow cytometry,respectively. The abilities of adhesion and invasion were detected by cell adhesion assay and cell invasion assay using Transwell chambers.RESULTS:The expression of MMP-2 was inhibited and the inhibitory effects of different sequence varied. pGPU6-1 group had the most efficient inhibitory effect,followed by pGPU6-2 and pGPU6-3 groups.Invasiveness and adhesion were more significantly reduced in pGPU6-1,pGPU6-2 and pGPU6-3 groups as compared with pGPU6 (-) and blank control groups. However,no difference concerning cell proliferation and apoptosis was observed after transfection between experiment groups and control groups.CONCLUSION:RNAi against MMP-2 successfully inhibited the mRNA and protein expression of MMP-2 in the pancreatic cancer cell line,BxPC-3,leading to a potent suppression of tumor cell adhesion and invasion without affecting cell proliferation and apoptosis. These findings suggest that the RNAi approach towards MMP-2 may be an effective therapeutic strategy for the clinical management of pancreatic tumor.

Relationship between matrix metalloproteinase-2 mRNA expression and clinicopathological and urokinase-type plasminogen activator system parameters and prognosis in human gastric cancer

AIM: To investigate the relationship between matrix metalloproteinase-2 (MMP-2) mRNA expression and clinicopathologic and urokinase-type plasminogen activator (uPA) system parameter and prognosis in human gastric cancer. METHODS: Expression of MMP-2 mRNA, uPA, and uPA-R mRNA in tumor tissues and ≥5 cm adjacent normal tissues from 67 cases of gastric cancer was studied using RT-PCR and Northern blot respectively.Survival analyses were done using the Kaplan-Meier method. RESULTS: The expression rates of MMP-2 mRNA,uPA and uPA-R mRNA in tumor tissues (31%,41%,and 51%, respectively) were significantly higher than those in ≥5 cm adjacent tissues (19%, 11%, and 9%; X2=4.59,43.58, and 53.24 respectively, P<0.05,0.0001,and 0.0001, respectively). Expression of MMP-2 mRNA was significantly correlated with lymph node metastasis (metastasis: 61.9%, no metastasis: 39.1%, X2= 7.61, P<0.05),Lauren's classification of diffuse/mixed types:54.2%,intestinal type: 26.3%,X2 = 4.25, P<0.05, expression of uPA and uPA-R mRNA (uPA+: 55.1%, uPA-: 22.2% and uPA-R+: 54.9%, uPA-R-: 18.8%, X2=5.72 and 6.40 respectively, P<0.05).Kaplan-Meier survival analysis of MMP-2 mRNA expression did not show significant difference in all 67 cases, but revealed an association of the expression of MMP-2 mRNA, uPA, and uPA-R mRNA with worse prognosis (P= 0.0083, 0.0160, and 0.0094, respectively). CONCLUSION: MMP-2 may play an important role in the development of invasion and metastasis of gastric cancer.

Correlation of RECK with Matrix Metalloproteinase-2 in Regulation of Trophoblast Invasion of Early Pregnancy

To study the role of the reversion-inducing-cysteine-rich protein with Kazal motifs （RECK） gene and matrix metalloproteinase-2 （MMP-2） in the regulation of trophoblast invasion of early pregnancy. Immunohistochemistry, Western blot and gelatin zymography were used to detect the RECK protein expression localization, expression level and MMP-2 activation level in the placental tissues harvested from 52 normal pregnant women （27 in the early pregnancy, 25 in the term pregnancy）. Immunohistochemistry showed that RECK expression was found both in villous tissues of early pregnancy group and term pregnancy group and was mainly observed in cell membrane and cytoplasm of cytotrophoblasts and syneytiotrophoblasts. RECK expression increased with gestational time. RECK expression of early pregnancy group was significantly lower than that of term pregnancy group （P〈0.05）. RECK expression was significantly lower in cellular column （CC） with invasion ability. Western blot showed that the RECK protein expression in early pregnancy group was significantly lower than that in term pregnancy （P〈0.05）. The optical density values of RECK protein expression in early pregnancy group and term pregnancy group were 1.35±0.14 and 2.68±0.26, respectively, while MMP-2 activation ratio was contrary to RECK protein expression and decreased with the gestation time （P〈0.01）. The MMP-2 activation ratios of early pregnancy group and term pregnancy group were 0.46±0.05 and 0.10±0.02, respectively. The expression of the tumor inhibitory gene RECK was positively related with the invasion ability of trophoblasts, while the invasion gene MMP-2 was negatively related with the ability. The interaction between RECK and MMP-2 may play an important role in the regulation of the trophoblast invasion in early pregnancy.

Distribution and relative activity of matrix metalloproteinase-2 in human coronal dentin

The presence of matrix metalloproteinase-2 （MMP-2） in dentin has been reported, but its distribution and activity level in mature human coronal dentin are not well understood. The purpose of this study was to determine the MMP-2 distribution and relative activity in demineralized dentin. Crowns of twenty eight human molars were sectioned into inner （ID）, middle （MD）, and outer dentin （OD） regions and demineralized. MMP-2 was extracted with 0.33 mol·L-1 EDTA/2 mol·L-1 guanidine-HCl, pH 7.4, and MMP-2 concentration was estimated with enzyme-linked immunoabsorbant assay （ELISA）. Further characterization was accomplished by Western blotting analysis and gelatin zymography. The mean concentrations of MMP-2 per mg dentin protein in the dentin regions were significantly different （P=0.043）： 0.9 ng （ID）, 0.4 ng （MD）, and 2.2 ng （OD）, respectively. The pattern of MMP-2 concentration was OD〉ID〉MD. Western blotting analysis detected -66 and -72 kDa immunopositive proteins corresponding to pro- and mature MMP-2, respectively, in the ID and MD, and a -66 kDa protein in the OD. Gelatinolytic activity consistent with MMP-2 was detected in all regions. Interestingly, the pattern of levels of Western blot immunodetection and gelatinolytic activity was MD〉ID〉OD. The eoneentration of MMP-2 in human coronal dentin was highest in the region of dentin that contains the dentinoenamel junction and least in the middle region of dentin. However, levels of Western blot immunodetection and gelatinolytic activity did not correlate with the estimated regional concentrations of MMP-2, potentially indicating region specific protein interactions.

Vascular Endothelial Growth Factorl65-regulated Nasopharyngeal Carcinoma Cell Lines Invasion and Migration Involve Expression and Activation of Matrix Metalloproteinase-2

The effect of vascular endothelial growth factor （VEGF） overexpression on matrix metalloproteinase-2 （MMP-2） in nasopharyngeal carcinoma （NPC） cells in vitro and the possible mechanism involved were investigated, and the correlation between the expression of VEGF and MMP-2 in NPC evaluated. The NPC cells were transfected with PAd-trackVEGF165 plasmid. The expression levels of VEGF and MMP-2 mRNA and protein in NPC cells were detected by semi-quantitative RT-PCR and Western blot respectively. It was found that the expression of VEGF and MMP-2 mRNA and protein was significantly increased in NPC cells after transfection of VEGF 165. It was concluded that the expression of VEGF was correlated to the in vitro invasion of NPC cells, and the induction of MMP-2 by VEGF was a key process of NPC cell invasion.

Expressions of cyclooxygenase-2 and matrix metalloproteinase-9 in cervical carcinoma and their clinical significance

Objective: To investigate the expressions of cyclooxygenase (COX)-2 and matrix metalloproteinase (MMP)-9 in cervical carcinoma and their clinical significance. Methods: Immunohistochemistry SP method was used to detect the expres- sions of COX-2 and MMP-9 in 72 cases of invasive carcinoma of cervix (ICC) and 16 cases of normal cervical epithelium remote from tumor (NCE). The relationships between the expressions of COX-2, MMP-9 in ICC and some characteristics relating to clinical pathology of cervical carcinoma such as histological grading, lymph node metastasis, stromal invasion and FIGO stage were analyzed statistically. Results: The rates of the positive expressions of COX-2 and MMP-9 in ICC were significantly higher than those in NCE. COX-2: 88.9% (64/72) in group ICC and 12.5% (2/16) in group NCE, P = 0.000; MMP-9: 94.4% (68/72) in group ICC and 43.8% (7/16) in group NCE, P = 0.000. The expression of COX-2 was positively correlated with lymph node metastasis (r = 0.296, P = 0.012) and stromal invasion (r = 0.257, P = 0.029). The expression of MMP-9 was positively correlated with FIGO stage (r = 0.329, P = 0.005) and histological grading (r = 0.351, P = 0.003). The expression of COX-2 was positively correlated with the expression of MMP-9 in ICC (r = 0.297, P = 0.011). Conclusion: The overexpressions of COX-2 and MMP-9 were closely related to the invasion and growth of cervical carcinoma. The tissue with the overexpression of COX-2 had strong invasion ability. COX-2 and MMP-9 had synergistic effect on proliferation, invasion and metastasis of cancer cells. Detecting the coexpression of COX-2 and MMP-9 may be of value in further understanding the biological behavior and predicting the prognosis of cervical carcinoma.

Rosiglitazone uppresses lipopolysaccharide-induced matrix metalloproteinase-2 activity in rat aortic endothelial cells via rasMEK1/2 signaling

ix metalloproteinase(MMPs) plays a key role in the pathogenesis of chronic inflammatory disease,such as atherosclerosis.Among MMPs,MMP-2 is regarded as a major proteinase in atherosclerotic plaque lesions.Peroxisome proliferator activated receptor-gamma(PPARg) ameliorates oxidative stress and the inflammatory response.The aim of the present study was to evaluate the effect of Rosiglitazone on Lipopolysaccharide(LPS)-induced MMP-2 activation as well as its possible mechanism.LPS-induced MMP-2 activity was inhibited by Rosiglitazone(PPARg agonist) in the rat aortic endothelial cells(RAEC).LPS-induced MMP-2 activation was diminished no matter exposure to NF-kB Activation Inhibitor II(JSH-23)or Ras inhibitor,farnesylthiosalicylic acid(FTS). Further study shows that LPS-induced activation of Phospho-Rho A and Phospho-MEKl/2 were significantly inhibited by Rosiglitazone.The activation of NF-kB p65 in the nuclear extract of cells was also significantly suppressed by Rosiglitazone, moreover,the expression of NF-κB p65 was partly activated by GW9662(PPARg antagonist).NF-kB DNA binding activity was also demolished by Rosiglitazone.In summary,our data showed that PPARg agonist,Rosiglitazone suppresses LPS-activated MMP-2 secretion via Ras-MEK1/2 signaling pathways and NF-kB activation.PPARg agonist and Ras-MEK1/2 pathway may be another potential therapeutic target for the disease induced by chronic inflammation.

Cryptotanshinone attenuates isoprenaline-induced cardiac fibrosis in the mouse associated with upregulation and activation of matrix metalloproteinase-2

Objectives The impairment of matrix metallopro- teinase-2(MMP-2)has been associated with the development of cardiac fibrosis.Although the Chinese herb Salvia miltior-rhiza has been widely used in patients with cardiovascular disorders,the mechanisms involved have not been elucidated. The purpose of the present study was to determine whether the administration of cryptotanshinone,an active ingredient of Salvia miltiorrhiza,could prevent the cardiac fibrosis induced by isoprenaline and to investigate the underlying mechanisms. Methods and Results Male C57BL/6 mice were submitted to receive daily injection of 0.9%saline,3 mg/kg isoprenaline, or isoprenaline plus 20 mg/kg cryptotanshinone by gastric gavage for 2 weeks.Herein,we demonstrate that cryptotanshinone can significantly ameliorate the isoprenaline-induced cardiac fibrosis,which was associated with marked up-regulation and activation of MMP-2 in ventricular myocardium. Additionally,we demonstrate that cryptotanshinone can dose-dependently upregulate and activate MMP-2 in cultured cardiac fibroblast.Moreover,incubation with cryptotanshinone also can prevent isoprenaline-induced downregulation and inactivation of MMP-2 in cultured cardiac fibroblast. Conclusions Taken together,our data suggest that cryptotanshinone may become a novel and potent antifibrotic agent. The present findings might further our understanding of the role of MMP-2 in cardiac fibrosis and antifibrotic mechanisms of cryptotanshinone.

Study on matrix metalloproteinase-2, 9 in peri-implant sulcular fluid

Objective: To study the expression of matrix metalloproteinases-2, 9 (MMP-2, MMP-9) of healthy implant and peri-implant sulcular fluid (PISF) by enzyme-linked immunosorbent assay (ELISA) method, and evaluate the level of MMP-2 and MMP-9 in sulcular fluid as an objective indicator of tissue inflammation around implants. Methods: A total of 40 implants were selected from 30 patients who were treated with dental implants and were divided into two groups: the inflammatory group and the healthy control group with 20 pieces respectively. ELISA double antibody sandwich method was used to detect the levels of MMP-2 and MMP-9 in PISF. Results: The MMP-2 and MMP-9 expressions were significantly different between the healthy implant group and the peri-implant group (p < .05). The concentration of MMP-2, MMP-9, and the amount of sulcular fluid in the inflammatory implant group were positively correlated with the clinical parameters (probing depth [PD], modified sulcus bleeding index [mSBI]). Conclusions: Under physiological conditions, the levels of MMP-2 and MMP-9 were low. When the periodontal tissue was stimulated by inflammation, the expression levels of MMP-2 and MMP-9 were increased, which could reflect the severity of inflammation. The increase levels of MMP-2 and MMP-9 in PISF could better reflect the health status of peri-implant tissues, which could be used as an objective indicator to assist in the diagnosis of peri-implant inflammation.

检测恶性腹水中VEGF、CD44v6和MMP-2、MMP-9的临床意义

背景及目的:恶性腹水中找到癌细胞有确诊意义,但阳性率很低。因此从腹水中寻找理想的癌性标记物已成为当前研究的主要课题。本课题拟通过检测腹水中血管内皮生长因子(vascularendothelialgrowthfactor,VEGF)、细胞粘附分子CD44v6、基质金属蛋白酶-2,-9(matrixmetalloproteinase-2,-9,MMP-2,-9)的水平并进行分析,以期为临床腹水诊治提供科学依据。方法:收集肝硬化腹水36例、结核性腹水8例、恶性腹水23例。采用ELISA法检测VEGF、CD44v6含量,用明胶酶谱法检测MMP-2,-9活性。结果:恶性腹水中VEGF、CD44v6含量分别为(640.74±264.81)ng/L、(89.22±38.20)μg/L,明显高于肝硬化腹水犤(67.05±51.91)ng/L,(44.79±18.02)μg/L犦和结核性腹水犤(88.25±24.12)ng/L,(50.25±12.57)μg/L犦(P<0.01)。肝硬化腹水、结核性腹水不能检出MMP-2,-9,而恶性腹水MMP-2检出率为87.0%,MMP-9检出率为78.3%。结论:恶性腹水中VEGF、CD44v6水平显著升高而MMP-2,-9阳性。联合检测腹水中VEGF、CD44v6、MMP-2,-9的水平可作为临床良、恶性腹水的鉴别诊断依据之一。