姜黄素通过上调miR-637抑制肾癌786-O细胞的增殖、迁移和侵袭

目的探讨姜黄素对肾癌786-O细胞的增殖、迁移和侵袭的影响及其可能的分子机制。方法体外培养人正常肾小管上皮细胞HK-2和肾癌786-O细胞, 将786-O细胞分为对照组、低剂量姜黄素组、中剂量姜黄素组、高剂量姜黄素组、miR-NC组、miR-637组、高剂量姜黄素+anti-miR-NC组、高剂量姜黄素+anti-miR-637组。分别进行实时荧光定量PCR(qRT-PCR)、MTT、Transwell和蛋白质印迹(Western blot)实验来检测各组的miR-637表达量、细胞活力、迁移侵袭能力和相关蛋白表达水平。结果与人正常肾小管上皮细胞HK-2比较, 肾癌786-O细胞中miR-637的表达水平降低(P<0.05)。与对照组比较, 中、高剂量姜黄素组786-O细胞的活力、迁移和侵袭细胞数、CyclinD1、MMP-2及MMP-9表达水平均显著降低, p21表达显著升高, miR-637的表达水平均显著升高(均P<0.05)。与miR-NC组比较, miR-637组的miR-637表达水平提高, miR-637组的细胞活力、迁移和侵袭细胞数、CyclinD1、MMP...     

芒柄花黄素调控miR-182/FOXO3轴对子宫内膜癌细胞迁移及侵袭的影响

目的 探讨芒柄花黄素(FM)对子宫内膜癌细胞迁移及侵袭的影响,以及其作用机制。方法 用0、12.5、25.0、50.0、100、200μmol/L FM处理人子宫内膜癌细胞株Ishikawa细胞48 h,流式细胞术检测细胞凋亡率;取生长状态良好的Ishikawa细胞,分别用25.0、50.0、100μmol/L FM处理,并命名为FM-L组、FM-M组、FM-H组,另取未处理的Ishikawa细胞作为对照组;利用细胞转染技术对处于对数生长期的Ishikawa细胞进行转染,分为miR-182 mimics组、miR-NC组、miR-182 inhibitor组、miR-NC inhibitor组、pc DNA-FOXO3组、pc DNA组,将miR-NC、miR-182mimics转染Ishikawa细胞后再用100μmol/L FM处理,记为FM+miR-NC组、FM+miR-182mimics组;Transwell测定细胞迁移及侵袭;Western Blot检测细胞中基质金属蛋白酶-2(MMP-2)、基质金属蛋白酶-9(MMP-9)、叉头转录因子O亚型3(FOXO3)蛋白表达;RT-...     

CCL21/CCR7对T24细胞生物学行为的影响

目的研究CCL21/CCR7对T24细胞迁移、侵袭、增殖以及抗凋亡能力的影响。方法采用不同浓度的CCL21(0、50、100、200ng/ml)作用T24细胞,用Transwell实验研究CCL21对T24细胞的迁移与侵袭能力;用MTT法研究了CCL21对T24细胞的增殖能力的影响,用FCM研究了CCL21对T24细胞抵抗ADM诱导的细胞凋亡,同时用Western blot检测了侵袭相关蛋白MMP-2与MMP-9表达及凋亡相关蛋白Bcl-2与Bax的表达。结果 CCL21作用于T24细胞能增强其迁移和侵袭能力,增强效果依赖于CCL21的浓度,CCL21能增加MMP-2与MMP-9的表达;CCL21还能促进T24细胞的增殖与抗凋亡能力,并能上调Bcl-2蛋白的表达,而抑制Bax蛋白的表达。结论 CCL21/CCR7增强了T24细胞的迁移与侵袭能力,促进了T24细胞的增殖与抗凋亡。     

lncRNA SNHG8通过靶向miR-335-5p调控膀胱癌T24细胞的增殖、迁移和侵袭

目的探讨lncRNA SNHG8对膀胱癌T24细胞增殖、迁移和侵袭的影响以及其分子作用机制。方法选取本院2017年1月至2019年1月收治的30例膀胱癌患者的癌组织及相应的癌旁组织;将膀胱癌T24细胞分为si-NC组、si-SNHG8组、miR-NC组、pcDNA组、pcDNA-SNHG8组、miR-335-5p组、si-SNHG8+anti-miR-NC组及si-SNHG8+anti-miR-335-5p组。对各组细胞采用qRT-PCR、MTT、Transwell、Western blot和双荧光素酶报告实验等检测并进行比较分析。结果与癌旁组织比较, 膀胱癌组织中SNHG8的表达水平升高, miR-335-5p表达水平下降(均P<0.001)。抑制lncRNA SNHG8或过表达miR-335-5p后, T24细胞的活力、迁移和侵袭能力显著下降, CyclinD1、MMP-2和MMP-9蛋白表达水平降低, p21蛋白表达水平升高(均P<0.001)。lncRNA SNHG8靶向调控miR-335-5p的表达水平。与si-SNHG8+anti-miR-NC组比较, si-S...     

异丙酚联合缺血预处理对大鼠肺缺血再灌注损伤时细胞凋亡的影响

目的 探讨异丙酚联合缺血预处理对大鼠肺缺血苒灌注损伤时细胞凋亡的影响.方法 雄性SD大鼠50只,200～250 g,随机分为5组(n=10):假手术组(S组)、缺血再灌注组(IR组)、异丙酚组(P组)、缺血预处理组(IP组)和异丙酚+缺血预处理组(P+IP组).阻断右肺门1 h后再灌注2 h制备大鼠单肺原位热缺血再灌注模型,P组夹闭右肺门前30 min持续静脉输注异丙酚30 mg·kg-1·h-1;IP组夹闭右肺门前先进行夹闭5 min,再灌注5 min,反复3次的缺血预处理;P+IP组夹闭肺门前30 min静脉输注异丙酚30 mg·kg-1·h-1和缺血预处理.于再灌注2 h时处死大鼠,取右肺下叶肺组织,光镜下观察肺组织病理学结果 ,计算肺损伤定量评价指标(IQA);检测肺凋亡细胞,计算肺细胞凋亡指数(AI);采用免疫组化法检测Bcl-2、Bax蛋白表达;IQA、Bcl-2/Bax蛋白比值与AI作直线相关分析.结果 与S组比较,IR组、P组、IP组和P+IP组再灌注后IQA、AI均明显升高,IR组Bcl-2、Bax蛋白表达上调,Bcl-2/Bax蛋白比值降低(P<0.01);与IR组比较,P组、IP组和P+IP组IQA、AI均明显降低,Bcl-2蛋白表达水平、Bcl-2/Bax蛋白比值升高,IP组、P+IP组Bax蛋白表达下调(P<0.01),P组差异无统计学意义(P>0.05);与P组和IP组比较,P+IP组IQA及AI降低,Bcl-2/Bax蛋白比值升高(P<0.05);IQA与AI呈正相关(r=0.951,P<0.01);AI与Bcl-2/Bax蛋白比值呈负相关(r=-0.851,P<0.01).结论 异丙酚联合缺血预处理可通过调节Bcl-2和Bax蛋白的表达抑制细胞凋亡,减轻肺缺血再灌注损伤.     

CircDONSON靶向miR-4458对肝癌Huh-7细胞增殖、迁移及侵袭的影响

目的:探讨CircDONSON对肝癌Huh-7细胞增殖、迁移及侵袭的影响及其可能的作用机制。方法:qRT-PCR法检测肝癌组织中CircDONSON、miR-4458表达量;肝癌细胞Huh-7分为si-NC组、si-CircDONSON组、miR-NC组、miR-4458组、si-CircDONSON+anti-miR-NC组、si-CircDONSON+anti-miR-4458组;CCK-8法、平板克隆形成实验、划痕实验与Transwell实验分别检测细胞增殖、克隆形成、迁移及侵袭情况;双荧光素酶报告实验检测miR-4458与CircDONSON靶向的关系;Western blot检测MMP-2、MMP-9蛋白表达量。结果:肝癌组织中CircDONSON表达量较癌旁组织升高(4.82±0.38比1.00±0.12,P<0.05),miR-4458表达量较癌旁组织降低(0.34±0.04比1.00±0.07,P<0.05);抑制si-CircDONSON或过表达miR-4458降低肝癌细胞活力、划痕愈合率和MMP-2、MMP-9蛋白水平(P<0.05)减少,细胞克隆...     

PI3K/Akt信号通路在异丙酚后处理减轻大鼠心肌细胞缺氧复氧损伤中的作用

目的 评价磷脂酰肌醇3-激酶/丝氨酸-苏氨酸蛋白激酶(PI3K/Akt)信号通路在异丙酚后处理减轻大鼠心肌细胞缺氧复氧损伤中的作用.方法 体外培养SD乳鼠心肌细胞,接种于96孔板(细胞密度1×105/ml,200 μl/孔)或6孔板(细胞密度5×105/ml,2 ml/孔)中,采用随机数字表法,将细胞随机分为4组(n=24):常规培养组(C组)细胞常规培养6h;缺氧复氧组(H/R组)细胞行缺氧2h,复氧4h;缺氧复氧+异丙酚组(H/R+P组)于缺氧结束时行异丙酚(终浓度50 μmol/L)后处理;H/R+异丙酚+ PI3K抑制剂组(H/R+ P+W组)于缺氧结束时加入PI3K抑制剂渥曼青霉素(终浓度100nmol/L)和异丙酚(终浓度为50μmol/L).复氧结束时,采用MTT法测定细胞活力,应用生化自动分析仪测定培养液LDH活性;流式细胞仪检测细胞凋亡情况;Western blot法检测心肌细胞磷酸化Akt(p-Akt)、Bcl-2及Bax表达水平.结果 与C组相比,H/R组细胞活力降低,LDH活性和细胞凋亡率升高,p-Akt和Bax表达上调,Bcl-2表达下调,Bcl-2/Bax降低(P＜0.05).与H/R组比较,H/R+P组细胞活力升高,LDH活性和细胞凋亡率降低,p-Akt和Bcl-2表达上调,Bax表达下调,Bcl-2/Bax升高(P＜0.05).与H/R+P组比较,H/R+ P+W组细胞活力降低,LDH活性和细胞凋亡率升高,p-Akt和Bcl-2表达下调,Bcl-2/Bax降低(P＜0.05).结论 异丙酚后处理减轻心肌细胞缺氧复氧损伤的机制与激活PI3K/Akt信号通路有关.     

长链非编码RNA氧化应激反应丝氨酸丰富1反义RNA 1对肾癌细胞增殖、迁移及侵袭的影响

目的探讨长链非编码RNA氧化应激反应丝氨酸丰富1反义RNA 1(lncRNA OSER1-AS1)对肾癌ACHN细胞的增殖、迁移、侵袭的影响及其对微小RNA(microRNA,miR)-612的调控作用。方法2017年1月至2020年3月,采用实时定量反转录聚合酶链反应(RT-qPCR)法检测肾癌组织、癌旁组织中OSER1-AS1、miR-612的表达量;体外培养肾癌细胞ACHN,分别将OSER1-AS1小分子干扰RNA(si-OSER1-AS1)及其阴性对照(si-NC)、miR-612寡核苷酸模拟物(miR-612 mimics)及阴性对照mimic NC序列(miR-NC)、si-OSER1-AS1与miR-612特异性寡核苷酸抑制剂的阴性对照(anti-miR-NC)、si-OSER1-AS1与miR-612特异性寡核苷酸抑制剂(anti-miR-612)转染至ACHN细胞;采用RT-qPCR法检测细胞中OSER1-AS1、miR-612的表达量;采用噻唑蓝(MTT)、Transwell小室实验分别检测细胞增殖、迁移及侵袭能力;双荧光素酶报告实验检测OSER1-AS1、miR-612的靶向关系;蛋白质印迹法(Western blot)检测细胞周期蛋白1(Cyclin D1)、基质金属蛋白酶(MMP)-2、MMP-9、p21蛋白表达量。两组间比较采用独立样本t检验,多组间比较采用单因素方差分析。结果肾癌组织OSER1-AS1的表达水平(1.00±0.08比3.37±0.28)高于癌旁组织(t=52.113,P<0.05),miR-612的表达水平(1.00±0.06比0.47±0.04)低于癌旁组织(t=47.062,P<0.05);si-OSER1-AS1组细胞活力(0.66±0.05比0.31±0.03)与Cyclin D1(0.63±0.05比0.25±0.02)、MMP-2(0.82±0.07比0.35±0.03)、MMP-9(0.76±0.06比0.28±0.03)蛋白水平低于si-NC组(t=18.007、21.169、18.514、21.466,P<0.05),si-OSER1-AS1组迁移细胞数[(115.56±9.52)个比(55.99±5.59)个]、侵袭细胞数[(93.41±6.09)个比(46.05±4.35)个]低于si-NC组(t=16.188、18.984,P<0.05),si-OSER1-AS1组p21蛋白水平(0.15±0.02比0.57±0.05)高于si-NC组(t=23.398,P<0.05);miR-612组细胞活力(0.68±0.05比0.39±0.03)与Cyclin D1(0.66±0.05比0.28±0.03)、MMP-2(0.85±0.06比0.46±0.03)、MMP-9(0.78±0.05比0.34±0.02)蛋白水平低于miR-NC组(t=14.920、19.551、17.441、24.512,P<0.05),miR-612组迁移细胞数[(118.76±9.87)个比(64.39±4.65)个]、侵袭细胞数[(99.65±9.12)个比(53.57±3.67)个]低于miR-NC组(t=14.950、14.062,P<0.05),miR-612组p21蛋白水平(0.14±0.02比0.51±0.04)高于miR-NC组(t=24.820,P<0.05);双荧光素酶报告实验证实OSER1-AS1可靶向结合miR-612;抑制miR-612表达可明显逆转干扰OSER1-AS1对细胞增殖、迁移及侵袭的作用。结论干扰OSER1-AS1可通过上调miR-612的表达从而抑制肾癌ACHN细胞增殖、迁移及侵袭。     

SNHG20调控miR-495对甲状腺癌生物学行为的影响

目的:探讨长链非编码RNA(lncRNA)核内小RNA宿主基因20(SNHG20)调控微小核糖核酸-495(miR-495)对甲状腺癌生物学行为的影响。方法:qRT-PCR法检测SW579、TPC-1、Nthy-ori3-1细胞中SNHG20、miR-495的表达;将SW579细胞分为CON组、si-NC组、si-SNHG20组、si-SNHG20+miR-NC组、si-SNHG20+miR-495 inhibitor组。检测细胞增殖、凋亡、周期和侵袭情况,细胞MMP-2、E-Cadherin、N-Cadherin蛋白表达情况,验证SNHG20和miR-495的关系。结果:与正常甲状腺上皮细胞相比,甲状腺癌细胞株SNHG20表达升高,miR-495表达降低(P<0.05),且SW579细胞SNHG20表达最高,miR-495表达最低,因此后续使用SW579细胞进行转染实验。SNHG20低表达会降低SW579细胞中SNHG20表达、存活率、侵袭个数以及MMP-2和N-Cadherin蛋白表达,升高细胞中miR-495表达、凋亡率、G0/G1期细胞比例以及E-Cadherin蛋白表达(P<0.05);miR-495抑制剂可减弱SNHG20低表达对SW579细胞增殖、侵袭的抑制作用,SNHG20靶向调控miR-495表达(P<0.05)。结论:低表达SNHG20可通过靶向调节miR-495抑制甲状腺癌细胞恶性生物学行为。     

LncRNA MIR205HG靶向miR-299-3p调控肺癌细胞增殖、凋亡的机制研究

目的 探讨长链非编码RNA MIR205HG(LncRNA MIR205HG)与微小RNA-299-3p(miR-299-3p)的相互关系及其影响肺癌细胞增殖、凋亡的分子机制。方法 检测肺癌组织和细胞中MIR205HG、miR-299-3p的表达。H1299细胞分为si-MIR205HG组、si-NC组、miR-299-3p组、miR-NC组、si-MIR205HG+anti-miR-299-3p组、si-MIR205HG+anti-miR-NC组。MIR205HG与miR-299-3p的相互作用；检测增殖、凋亡及蛋白表达。结果 肺癌组织和细胞系中MIR205HG表达水平升高(P<0.05),miR-299-3p表达水平降低(P<0.05)。MIR205HG能与miR-299-3p特异性结合，可调控miR-299-3p的表达。干扰MIR205HG表达或miR-299-3p过表达后可降低肺癌细胞增殖能力，促进细胞凋亡，且CyclinD1、p21表达降低，Bcl-2、Bax表达增高(P<0.05)。抑制miR-299-3p可逆转干扰MIR205HG表达对肺癌细胞增殖及凋亡...     

Priestley, the furious free thinker of the enlightenment, and Scheele, the taciturn apothecary of Uppsala

'The more elaborate our means of communication, the less we communicate'.
Joseph Priestley     

Comparative Anatomy of the Subsynovial Connective Tissue in the Carpal Tunnel of the Rat, Rabbit, Dog, Baboon, and Human

The tenosynovium in the human carpal tunnel is connected to the flexor tendons and the median nerve by the subsynovial connective tissue (SSCT). The most common histological finding in carpal tunnel syndrome (CTS), a compression neuropathy of the median nerve, is noninflammatory fibrosis of the SSCT. The relationship, if any, between the fibrosis and nerve pathology is unknown, although some have speculated that a change in the SSCT volume or stiffness might be the source of the compression. Yet, while animal models have been used to study the physiology of nerve compression, so far none have been used to study the relationship of the SSCT pathology to the neurophysiological abnormalities. The purpose of this study was to identify animal models that might be appropriate to study the interaction of SSCT and nerve function in the development of CTS. The front paws of a rat, rabbit, dog, and baboon were dissected. The carpal tunnel anatomy and SSCT of these animals were also examined by light and scanning microscopy and compared to the relevant human anatomy and ultrastructure. The carpal tunnel anatomy and contents of the baboon and rabbit are similar to humans. The canine carpal tunnel lacks the superficial flexor tendons and the rat carpal tunnel is very small. The human, baboon, and rabbit specimens had very similar organization of the SSCT, and content of the carpal canal. We conclude that, while both the baboon and rabbit would be good animal models to study the relationship of the SSCT to CTS, the rabbit is likely to be more practical, in terms of cost and animal care concerns.     

Effect of castration, DES, flutamide, and the 5 alpha-reductase inhibitor, MK-906, on the growth of the Dunning rat prostatic carcinoma, R-3327.

Male rats bearing implants of the Dunning rat prostatic carcinoma, R-3327, were used in a 42-day study to determine the effect of castration or orally administered flutamide (FL), DES (diethylstilbestrol) or the 5 alpha-reductase inhibitor, MK-906, on the growth of this androgen-responsive cancer. The rate of growth and final weights of the tumor and the ventral prostate (VP) were all reduced (P less than 0.05) by castration. Flutamide (25 mg/kg/day) significantly decreased tumor and VP weights in intact rats and castrates given 100 micrograms/day (SC) of testosterone propionate (TP) or dihydrotestosterone propionate (DHTP). It also significantly retarded tumor growth rate in TP- or DHTP-treated castrates and was marginally effective in intact animals. DES (100 micrograms/kg/day) reduced (P less than 0.05) tumor and VP weights of intact rats but did not significantly affect tumor growth rate or weight in castrates given TP or DHTP. These results indicated that the effect of DES on tumor growth is caused by its inhibition of the secretion or release of the gonadotropins necessary for testicular androgen production. MK-906 (25 mg/kg/day) affected neither the gross nor the histomorphology of the tumor in intact rats or castrates given TP or DHTP. Further, it caused no histological changes in the testes of intact rats. It did, however, significantly reduce VP weight in intact animals and TP-treated castrates but not in those given DHTP. This illustrates that the anti-androgenicity of MK-906 stems from its inhibition of DHT formation. The failure of MK-906 to influence tumor growth in the TP-treated castrates strongly suggests that the R-3327 tumor can respond to testosterone directly. If that is true, then its growth is unlikely to be affected by a pure 5 alpha-reductase inhibitor such as MK-906. In ancillary experiments, tumors from MK-906-treated animals were found to have reduced levels of DHT and, when assayed in vitro, to have a reduced capacity to convert [3H]-T to [3H]-DHT.     

Prostate cancer, the urologist, the patient, and treatment

Prostate cancer, particularly advanced prostate cancer, should be considered as a chronic disease that requires multidisciplinary management. Each health professional involved the urologist of course, but also the other therapists, especially the general practitioner and nurse, should place their activities within a therapeutic synergy. The onco-psychological dimension should not be underestimated. In particular, information about the disease, its course and the therapeutic solutions proposed should be given gradually and, above all, should be regularly recapitulated. Patients are more accepting of treatments when they know about them. The point of view of the patient with prostate cancer is approached here in two ways; firstly through the comments of a psychologist on the history of a patient with prostate cancer and the dialogue with an experienced urologist. The treatment should not only be understood, but also accepted and requested by the patient. In this chronic disease, the patient often feels the treatment to be a bond between the therapist and himself. A survey based on an auto-questionnaire was also conducted among 275 patients and 50 urologists in order to investigate these relationships. From the results of this survey, in the case of hormone treatment, a quarterly rhythm for LHRH agonist injections appears to be perfectly matched to patient requirements. Monthly injections involve too great an intrusion by the treatment into the life of a cancer patient, in contrast injections which are spaced at greater than quarterly intervals could be seen to much as desertion and also lead to forgetfulness and neglect.