PHLPP1在卵巢浆液性囊腺癌的表达及对预后的影响

目的:检测PHLPP1蛋白在卵巢浆液性囊腺癌中的表达,探讨PHLPP1的表达对初治卵巢浆液性囊腺癌耐药及预后的影响。方法:收集整理2007年1月至2009年8月资料完整且病理确诊的卵巢浆液性囊腺瘤26例和卵巢浆液性囊腺癌93例的临床资料,对所有患者术后石蜡组织行免疫组织化学检测PHLPP1与磷酸化丝/苏氨酸蛋白激酶B(p-AKT)的表达。通过Kaplan-Meier法进行生存分析,并对p-AKT与PHLPP1的表达相关性进行分析,COX回归模型对影响卵巢浆液性囊腺癌预后的临床及病理因素进行分析。结果:PHLPP1在卵巢浆液性囊腺瘤的高表达率为61.5%,在浆液性囊腺癌高表达率为41.9%,两组比较差异有统计学意义(χ~2=42.10,P0.001)。PHLPP1与p-AKT表达呈负相关(r=-0.513,P0.001)。PHLPP1表达与卵巢浆液性囊腺癌恶性程度、淋巴结转移、远处转移、铂类耐药及分期相关,PHLPP1无或低表达及p-AKT高表达、淋巴结转移、远处转移是影响其预后的独立危险因素。卵巢浆液性囊腺癌PHLPP1高表达组中位生存期(MST)为48.0±7.6月,平均复发时间为10.2±3.3月;无或低表达组MST 34.0±4.0月,平均复发时间6.0±2.9月,高表达组均高于无或低表达组(P0.05)。PHLPP1高表达组5年生存率为25.0%,无或低表达组5年生存率为16.1%。p-AKT高表达组5年生存率为9.4%,而无或低表达组为23.3%。结论:PHLPP1在卵巢浆液性囊腺瘤中的高表达率显著高于浆液性囊腺癌,PHLPP1表达与卵巢浆液性囊腺癌患者耐药及预后密切相关。PHLPP1高表达者预后明显好于无或低表达者,PHLPP1缺乏是浆液性囊腺癌的独立危险因素。     

脐灸治疗脊髓损伤后神经源性膀胱30例临床观察

目的观察脐灸治疗脊髓损伤后神经源性膀胱的临床疗效。方法将60例脊髓损伤后神经源性膀胱患者随机分为治疗组和对照组各30例,两组患者均给予常规康复训练及脊髓损伤护理,治疗组加用脐灸治疗,将茯苓、苍术、炮附片、干姜、炒白芍等中药制成脐灸粉施灸于神阙穴,2或3天1次,3次为1个疗程,治疗2个月。分别于治疗前后各1周B超监测两组患者平均膀胱容量、残余尿量、日均单次排尿量,并进行生存质量评分和下尿路症状(LUTS)评分。结果治疗后两组患者的膀胱容量、残余尿量、日均单次排尿量及生存质量评分和LUTS评分均明显改善(P0.05),且治疗组各项目改善程度优于对照组(P0.05)。结论脐灸可以有效改善脊髓损伤后神经源性膀胱患者膀胱功能,并提高患者生活质量。     

卵巢癌腹水CD4+ CD25+调节性T细胞免疫抑制功能及机制的研究

  目的  研究卵巢癌患者腹水内CD4+ CD25+调节性T细胞（Treg）免疫抑制功能与患者临床病理特点的关系及其与患者初治及复发状态是否相关,并进一步探讨卵巢癌腹水内CD4+ CD25+调节性T细胞（Treg）发挥免疫调节性作用的具体机制。  方法  应用免疫磁珠分选法分选28例卵巢癌患者腹水内CD4+ CD25+调节性T细胞及CD4+ CD25- T细胞,采用羧基荧光素二醋酸盐琥珀酰亚胺酯（carboxyfluorescein succinimidylester,CFSE）标记CD4+ CD25- T细胞,CD4+ CD25+ Treg与自体CFSE标记CD4+ CD25- T细胞以1:1比例共培养,经流式检测CFSE表达及Modfit软件分析CD4+ CD25- T增殖指数（PI）,计算各标本内Treg对自体CD4+ CD25- T细胞增殖抑制率。应用中合性抗IL-10抗体及中合性抗TGF-β1抗体探究腹水内CD4+ CD25+ Treg介导免疫逃逸作用是否通过抑制性细胞因子IL-10或TGF-β1发挥作用。  结果  Ⅲ~Ⅳ期卵巢癌腹水内CD4+ CD25+ Treg对自体CD4+ CD25- T细胞增殖抑制率为（75.72±17.04）%,较Ⅰ~Ⅱ期Treg抑制率（59.61±16.97）%显著增强（P < 0.05）。复发卵巢癌患者腹水内Treg对自体CD4+ CD25- T细胞增殖抑制率为（85.89±7.07）%,较初治卵巢癌患者腹水Treg抑制率（52.82±8.18）%显著增强（P=0.000 1）。共培养体系内加入中合性抗IL-10抗体或/及中和性抗TGF-β1抗体,Treg对自体CD4+ CD25- T细胞增殖抑制率较对照组均显著降低（P < 0.05）。  结论  卵巢癌腹水内CD4+ CD25+ Treg介导免疫逃逸能力与肿瘤分期及复发、初治状态相关,且发挥免疫逃逸作用可能是与分泌抑制性细胞因子IL-10及TGF-β1有关。      

卵巢癌患者腹水及外周血CD4+ CD25+调节性T细胞含量及抑制功能的研究

  目的   研究卵巢癌患者腹水及外周血单个核细胞（PBMC）中CD4+ CD25+调节性T细胞（Treg）的表达差异、免疫调节功能及其含量与化疗、复发的关系,探究其在卵巢癌腹腔微环境中发挥免疫调节的具体作用及意义。   方法   采用流式细胞术分别检测27例卵巢癌患者腹水及28例卵巢癌患者PBMC中CD4+ CD25+/CD4+ T细胞百分比,并根据收集标本时患者临床特征进行分组,比较处于初治（PD）、化疗后（AC）及复发（RD）3个阶段卵巢癌患者腹水及PBMC中Treg含量的差异。应用免疫磁珠分选卵巢癌患者腹水及外周血Treg,与经羧基荧光素二醋酸盐琥珀酰亚胺酯（carboxyfluorescein succinimidylester,CFSE）标记的自CD4+CD25- T细胞按不同比例（0:1,1:1,1:2及1:4）共培养,检测Treg免疫抑制功能。   结果   卵巢癌患者腹水中CD4+ CD25+/CD4+ T细胞百分比（28.25±14.06）%较PBMC中（14.6±4.74）%显著增高（P < 0.000 1）。卵巢癌患者PD、AC及RD 3个阶段腹水及PBMC中Treg含量均显示为AC>RD>PD,且均有显著统计学意义（P < 0.000 1）。体外实验结果显示,卵巢癌患者腹水中CD4+ CD25+ Treg可有效抑制CFSE标记的自体CD4+ CD25- T细胞增殖,且抑制功能较外周血显著增强（P < 0.01）。   结论   卵巢癌腹腔微环境中存在CD4+ CD25+Treg,且其含量及免疫抑制功能较外周血PBMC显著增高,提示卵巢癌腹腔内更易发生免疫逃逸作用。卵巢癌患者化疗后及复发阶段腹水及PBMC中CD4+ CD25+ Treg含量均大于原发阶段,提示化疗可能促进卵巢癌患者体内Treg含量升高,而Treg升高可能参与促进肿瘤复发。       

原发性骶前肿瘤147例临床病理分析

回顾性分析1954～2004年收治的147例原发性骶前肿瘤患者的临床病理资料,结果显示,原发性骶前肿瘤的主要诊断方法包括直肠指诊、B超和CT检查;肿瘤病理类型复杂,147例共涉及不同病理类型30种,其中良、恶性分别为49.7%和50.3%;主要治疗手段为手术治疗,本组病例肿瘤完整切除者77例,部分切除者32例,二次手术者18例;78例骶前恶性肿瘤联合应用放、化疗或免疫治疗,综合治疗的疗效与原发肿瘤的组织学类型、肿瘤体积等因素有关;患者生存期3个月～14年,中位生存期2年,影响预后的主要因素为肿瘤的组织学类型、肿瘤的手术切除率及综合治疗的应用。     

沉默轴突导向蛋白4D基因对卵巢癌SKOV3细胞恶性生物学行为的影响

Objective To observe the effect of DNA Sema4D gene silencing by RNA interfering on the proliferation, migration and invasion of human ovarian cancer SKOV3 cells, and to study the effect of pshRNASema4D on the growth of SKOV3 cells in transplanted tumor in nude nice. Methods Recombinant plasmid pshRNASema4D-A, B and C were respectively transfected into SKOV3 cells by lipofetamine 2000, while cells transfected by plasmid vector pcDNA3.1 and cells untreated as control groups. RT-PCR was adopted to select the recombinant plasmid which showed the most optimal inhibition effect. RT-PCR and Western blotwere used to detected the mRNA and protein expression of Sema4D in SKOV3 cells tranfected for 24, 48 and 72 hours. MTT assay was used to investigate the proliferation of the SKOV3 cells after trasnsfection. Transwell cell migration and invasion assays were used to investigate the migration and invasion abilities of the SK0V3 cells after trasnsfection. Human ovarian cancer model was established in nude mice, and the nude mice were treated with pshRNASema4D-B once every 3 days for 3 weeks. The bulk of the transplanted tumor was measured. Results Three Sema4D-targeted short hairpin RNA (shRNA) A, B and C were successfully inserted into the plasmid vector pshRNA, and the coding sequences of the obtained shRNA were consistent with the designed fragment. The results indicated that both recombinant plasmid pshRNASema4D-A and B could effectively knock down the expression of Sema4D gene in human ovarian cancer cells, of which pshRNASema4D-B was the better choice, while no effect of pshRNASema4D-C was seen. RT-PCR results showed that the relative mRNA expression of Sema4D gene in SKOV3 cells transfected with pshRNA-Sema4D for 24, 48 and 72 hours were 0. 401 ±0.051, 0. 120 ±0.035 and 0.014 ±0. 015, respectively, which were significantly lower than that in SKOV3 cells transfected by empty vector and non-transfected cells at 72 hours after transfection. (0.521 ±0.019, 0.536 ±0.040,respectively, P<0.05). The Westen blot analysis manifested that the relative expression of Sema4D protein of SKOV3 cells transfected by pshRNASema4D for 24, 48 and 72 hours were 0. 196 ± 0. 023, 0. 074 ± 0. 015 and 0. 040 ± 0.014, respectively, which were significantly lower than that in SKOV3 cells transfected by empty vector and non-transfected cells at 72 hours after transfection. (0. 275 ± 0. 009, 0. 282 ± 0. 015, respectively, P < 0. 05 ). Comparing with the empty vector-transfected and non-transfected cells, the proliferation, invasion and migration ability of SKOV3 cells transfected with pshRNA-Sema4D were obviously weakened. The pshRNASema4D-B significantly suppressed the growth of the SKOV3 cells-transplanted tumors in nude mice, and the IR( inhibitory rate ) of pshRNASema4D-B group was ( 61.0 ± 3.3 ) % ( P < 0.05). Conclusions Sema4D can be successfully silenced by RNA interfering in human ovarian cancer SKOV3 cells. Downregulation of Sema4D can inhibit the proliferation, migration and invasion of ovarian cancer cells. The pshRNASema4D can significantly suppress the growth in transplanted tumor of human ovary cancer in nude mice. Sema4D may become a candidate gene of gene therapy of human ovarian cancer.     

分子靶向治疗在妇科恶性肿瘤中的研究进展

分子靶向治疗（molecular targeted therapy,MTT）作为肿瘤治疗的新手段,通过特异性靶向肿瘤发生发展中起关键作用的分子或相关细胞的信号转导通路控制细胞基因表达,从而抑制或杀死癌细胞,基于对细胞分子生物学及信号转导机制,在分子水平上对肿瘤细胞进行直接或间接的精确打击。肿瘤分子靶向治疗因其具有疗效高、不良反应低的特点而备受瞩目,其出现为肿瘤的治疗开辟了新的领域和广阔的前景。本文就妇科恶性肿瘤中分子靶向治疗的研究进展进行综述。      

卵巢癌腹腔微环境中肿瘤相关巨噬细胞表型特点及其TLR表达谱的研究

  目的  探讨卵巢癌腹水微环境中肿瘤相关巨噬细胞(TAM)浸润情况及其表型特征和TAM的Tol样受体表达情况。  方法  应用免疫组织化学方法及流式细胞术检测卵巢癌及妇科良性病变腹水中CD68+TAM及其亚型的比例情况。应用Real-timePCR检测不同亚型TAM的Tol样受体的表达情况。  结果  23例卵巢癌腹水细胞CD68+TAM平均值为73.6个/HP, 20例良性病变平均计数为50.2个/HP。两组间CD68+TAM浸润差异无统计学意义(P > 0.05)。CD68+TAM在卵巢癌及对照组腹水中所占比例无差异, 分别为31.7%和18.8%(P > 0.05)。卵巢癌腹水中M2型TAM占全部巨噬细胞54.3%, 明显高于对照组10.0%(P < 0.05);对照组M1型TAM占优势, 为53.3%, 明显高于卵巢癌组的30.5%(P < 0.01)。不同亚型巨噬细胞Toll样受体表达差异无统计学意义。  结论  卵巢癌腹腔微环境中同时存在M1和M2型TAM, 但M2型TAM在卵巢癌中占优势, 而M1型TAM在良性病变中占优势。Toll样受体不能区分M1-TAM与M2-TAM。