目的:研究配对盒基因1(pax1)在子宫颈组织中的蛋白表达及其临床意义,探讨pax1蛋白与子宫颈癌预后的关系.方法:采用免疫组化法检测中日友好医院2011年8月至2017年10月收治的101例子宫颈癌、120例子宫颈上皮内瘤变(CIN)及20例正常子宫颈组织(对照组)中pax1蛋白表达水平,分析其子宫颈癌临床病理参数,采用受试者工作曲线(ROC)评估其诊断效能并随访,探讨其与子宫颈鳞癌(Scc)及子宫颈腺癌(AC)预后的关系.结果:①对照组的pax1蛋白表达水平(90.0％)高于CIN(73.3％)及子宫颈癌(38.6％),SCC患者pax1蛋白阳性表达率(20.9％)明显低于AC (70.0％)及特殊类型子宫颈癌(100.0％),差异有统计学意义(P＜0.05).②ROC曲线显示,pax1蛋白曲线下面积(AUC)为0.829,诊断子宫颈癌的灵敏度90.0％,特异度61.3％.③pax1蛋白阳性表达率与子宫颈癌患者的HPV感染、分化程度、组织类型及脉管浸润有关(P＜0.05).④Scc患者中pax1蛋白阳性组的生存时间明显高于阴性组(P =0.040),AC患者中pax1蛋白阳性组的生存时间与阴性组比较,差异无统计学意义(P=0.589).结论:pax1蛋白的表达缺失与CIN及子宫颈癌进展密切相关,其诊断CINⅡ及以上的效能较高.pax1蛋白的表达缺失与SCC的发生发展及预后有一定的关系,pax1蛋白有望成为监测SCC患者预后的有效指标.

目的:研究含缬氨酸蛋白p97在宫颈病变中的表达水平,探讨p97蛋白与宫颈癌发生发展的相关性,并分析其对于宫颈癌远处转移及预后评估的临床价值.方法:收集中日友好医院201 1年8月～2017年10月妇科手术标本共259例,其中宫颈癌101例、上皮内病变(CIN)138例(其中CIN1 48例、CIN2 50例、CIN3 40例)、正常宫颈20例.采用免疫组化法检测p97蛋白表达,采用单因素Kaplan-Meier法分析p97蛋白对宫颈癌总生存率(OS)的影响.结果:p97蛋白表达在正常宫颈组、CIN组、宫颈癌组3组间差异均有统计学意义(均P＜0.05),p97蛋白与HPV感染、病理分化程度、肿瘤直径、淋巴转移及SCC值相关(P＜0.05).K-M生存分析结果显示,p97蛋白高表达组的总生存率显著低于p97蛋白低表达组(生存中位数34.27个月vs.51.55个月,P＜0.01).结论:p97蛋白与宫颈癌的发生发展及远处转移密切相关,它有望成为检测宫颈癌病情进展及评估预后的生物标记物.

Curcumin, as a high effect and low toxicity anti-cancer drug and photosensitiser, has synergistic and complementary effects with photodynamic therapy (PDT). However, due to its unclear mechanism, PDT's application and efficacy were limited. Notch signaling pathway, which is highly correlates with carcinogenesis and development of cervical cancer, could be a potential therapeutic targets to improve the effectiveness of PDT. Therefore, in this study, we explored the effects of Notch signaling pathway in cervical cancer by curcumin mediated PDT with/without Notch receptor blocker (DAPT), and hope to elucidate its mechanism. Firstly, the effect on the proliferation of cervical cancer Me180 cells were detected with MTT assay, and apoptosis were detected with Annexin V-FITC/PI combined with flow cytometry. Secondly, after establishment of nude mice model, dividing the experimental animals into model group, curcumin PDT group, simple DAPT group, and curcumin-PDT+DAPT group, and analyzing tumor volume changes as well as HE staining in each group. mRNA and protein expression of gene Notch-1 and its downstream NF-κB and VEGF were observed with RT-PCR, immunohistochemical staining and Western-blot with/without inhibition of Notch signaling pathway by DAPT, both in vivo and in vitro experiments. We found both DAPT and curcumin-PDT can inhibit the proliferation and induce apoptosis of cervical cancer cell. The two have synergistic effect in vitro and in vivo. This effect can effectively block the conduction of Notch signaling pathway, which is associated with down-regulation of the expression of Notch1 and NF-κB. Notch signaling pathway could be one of the targets of curcumin-PDT photodynamic therapy.

目的:探索P16在宫颈上皮内瘤变Ⅱ级(CINⅡ)组织中的表达状态及其作为生物标记物对CINⅡ患者进行分流管理和预后判断的临床意义.方法:选取中日友好医院2008年1月～2017年12月宫颈锥切术后病理诊断为CINⅡ的组织学标本158例,应用免疫组织化学法检测P16的表达,取正常宫颈组织40例进行对照研究,并对CINⅡ患者进行术后随访统计.结果:CINⅡ患者宫颈组织中P16阳性表达率为55.7％,显著高于正常宫颈组织中P16阳性表达率(7.5％,P＜0.01).在CINⅡ患者中随机选取37例进行术后随访,P16阳性患者超薄细胞检测(LCT)阳性率72.72％,显著高于P16阴性患者LCT阳性率(15.79％,P＜0.05).P16阳性表达患者的HPV阳性率为75％,显著高于P16阴性表达患者的HPV阳性率(18％,P＜0.05).结论:P16在CINⅡ患者阳性表达提示病情进展风险更高,建议积极进行宫颈锥切诊断治疗,并加强术后随访;P16阴性患者建议暂时观察,避免不必要的手术创伤.

宫颈癌是妇科常见恶性肿瘤,在宫颈癌细胞也观察到了p97蛋白阳性表达升高.随着近年来对其深入研究,p97蛋白成为肿瘤治疗的新靶点,研究表明,含缬氨酸蛋白p97与多种恶性肿瘤的发生发展、远处转移及预后相关,p97蛋白参与众多生理病理过程,主要通过清除细胞内的异常蛋白质,控制蛋白质质量,维持正常细胞及肿瘤细胞存活[1,2].p97抑制剂在治疗妇科恶性肿瘤方面有其独特的应用.本文就p97蛋白宫颈癌的相关性研究进展进行综述,以期为宫颈癌的预防和治疗提供更多参考.

目的:探讨HPV E6/E7蛋白在宫颈癌早期筛查方面的临床应用价值.方法:收集2015年11月～2017年11月间中日友好医院妇产科宫颈病变门诊就诊和住院的400例患者,选取其中53例HPV16/18型阳性患者检测E6和E7蛋白的表达,分析其表达水平与组织病理学符合情况.结果:HPV E6蛋白在炎症、CIN 1、CIN 2-3和宫颈癌中的阳性表达率为0、37.5％、64.0％和84.6％;HPV E7蛋白分别为14.3％、37.5％、80.0％和92.3％.随着病变级别增加,HPV E6/E7蛋白的阳性表达率显著增加(P＜0.01).HPV E6、E7蛋白检测的灵敏度(71.1％、84.2％)均高于LCT(63.2％),阳性预测值(90.0％、88.9％)均高于HPV-DNA检测(71.7％).结论:HPV E6/E7蛋白可能成为高度鳞状上皮内病变及宫颈癌的早期检测标志物,其检测可能提高筛查的准确性.

Objective:To search the optimum parameters of curcumin as a photosensiti-zer of photodynamic therapy induced apoptosis of human cervical carcinoma Me180 cells by or-thogonal design. Methods:The selected photosensitizer curcumin concentration,laser dosage,ir-radiation time, incubation time four factors, according to the concentration of curcumin 2. 5μmol/L,5μmol/L,10μmol/L,50J/cm2 ,100J/cm2 ,200J/cm2 laser dose,irradiation time:90s, 180s,360s,incubation time:12h,24h,36h. 3 levels of each factor design and orthogonal design table,photodynamic therapy of curcumin by orthogonal design table as a photosensitizer,after treatment of cells,cell survival rate was measured by MTT method. The average marginal value of three levels of curcumin and laser dosage were estimated by two factor analysis of variance. Results:The order of influence factors of cell apoptosis induced by PDT is:curcumin dose >in-cubation time > the laser irradiation time >laser dosage. The level of various factors:Curcumin concentration:10μmol/L>5μmol/L>2. 5μmol/L;200J/cm2>100J/cm2>50J/cm2 laser dose;irradiation time:360s>180s>90s;incubation time:24h>12h>36h. Double factor variance analy-sis showed that curcumin concentration of 2. 5μmol/L,5μmol/L,10μmol/L among the three groups were statistically significant differences(P<0. 05);100J/cm2,200J/cm2 and 50J/cm2 group,there was a statistically significant difference (P<0. 05),100J/cm2 and 200J/cm2 but no statistically significant difference between the groups. Conclusions:Curcumin mediated photo-dynamic therapy induced apoptosis of cervical cancer cells with the best combination of curcu-min concentration of 5μmol/L,laser dose of 100J/cm2 ,irradiation time of 180s,incubation time of 24h.

Objective To explore the mechanism of notch signaling pathway in the treatment of cervical cancer with notch receptor blocker(DAPT).Methods After treatment of Me180 cells with different concentrations of DAPT combined with curcumin-photodynamic therapy (PDT),3-(4,5 Dimethylthiazlo-yl)-2,5 Dimethylthiazol-2-yl)-2,5 dipheny tetrazo lium bromide(MTT)assay was used to detect the effect on the proliferation of Me180 cells,annexin V-FITC/PI combined with flow cytometry was used to detect the effect on apoptosis,and real-time fluorescence quantitative PCR and Western blot were used to detect the mRNA and protein expressions of Notch1 gene respectively.Results Both DAPT(1.0 μmol/L)and curcumin-PDT groups could inhibit the proliferation and induce apoptosis of cervical cancer cell line Me180,and the synergistic effect of the two groups was significant (P＜0.05 or＜0.01).DAPT and curcumin-PDT group reduced the mRNA expression level of Notch1 in cervical cancer Me180 cells with inhibition ratio of 40.0％ and 32.3％ respectively.DAPT and curcumin-PDT group inhibited the protein expressions of Notch1,nuclear factor-kappa B(NF-kB),vascular endothelial growth factor (VEGF),and the synergistic effect of the two groups was statistically significant.Conclusions Both DAPT and curcumin-PDT may effectively block the conduction of Notch signaling pathway,which is associated with down-regulation of the expression of Notch1 and NF-kappa B.Notch signaling pathway may be one of the targets of curcumin photodynamic therapy.