CD44基因多态性与宫颈癌及非小细胞肺癌的相关性

背景与目的：CD44分子是众多肿瘤细胞的标志分子，其表达水平与肿瘤细胞的恶性程度有关。该研究探讨CD44基因中的单核苷酸多态性（single nucleotide polymorphism，SNP）位点与云南汉族人群宫颈癌和非小细胞肺癌（nonsmall cell lung cancer，NSCLC）易感性的相关性。方法：选取了CD44基因中的两个SNP位点rs13347和rs8193，采用TagMan基因分型的方法，分析这两个多态性位点在497例宫颈癌患者和500例健康对照个体以及483例NSCLC患者和471例健康对照个体中的分布特征，并分析CD44基因中的多态性位点与云南汉族人群宫颈癌和NSCLC的相关性。结果：rs13347和rs8193位点等位基因和基因型在宫颈癌组和对照组中的分布频率的差异无统计学意义（P>0.05）。而在NSCLC组和对照组的比较中：rs13347和rs8193位点等位基因在NSCLC组和对照组中的分布频率的差异有统计学意义（P=0.020和P=0.004）；这两个位点基因型在NSCLC组和对照组中分布频率的差异有统计学意义（P=0.027和P=0.020）；其中rs13347位点等位基因C在NSCLC组中的分布频率显著高于对照组，可能是NSCLC发生的风险因素（OR=1.250，95% CI：1.035～1.509），rs8193位点等位基因C在对照组中的分布频率显著高于NSCLC组，可能是NSCLC发生的保护性因素（OR=0.768，95%CI：0.641～0.921）。单倍型分析结果显示，rs13347C-rs8193T和rs13347T-rs8193C在NSCLC组和对照组中的分布频率差异有统计学意义（P=0.003和0.022）；该结果说明单倍型rs13347C-rs8193T可能是云南汉族人群NSCLC发生的风险性因素（OR=1.316，95%CI：1.096～1.579）。结论：CD44基因中的两个SNP位点rs13347和rs8193可能与云南汉族人群宫颈癌发病风险无关，而可能与云南汉族人群NSCLC具有相关性。     

Low-dose all-trans retinoic acid enhances cytotoxicity of cisplatin and 5-fluorouracil on CD44+ cancer stem cells

Cis-diamminedichloridoplatinum(II)(CDDP)-based combination chemotherapy is frequently used in gastrointestinal cancer. The synergistic mechanism of all-trans retinoic acid (ATRA), cisplatin (CDDP) and 5-fluorouracil (5-FU) in combination remains unclear. Despite their potent antitumor properties, resistance to CDDP and 5-FU develops frequently in tumors. To clarify this mechanism, we determined the sensitivity to each drug and their combination in two gastrointestinal cancer stem cells (CSCs) subpopulation.Here, we report the identification and separation of CD44⁺ cells from human gastric carcinoma (AGS) and human esophageal squamous cell carcinoma (KYSE-30) cancer cell lines by magnetic activated cell sorting (MACS). We allowed the CD44^± cells to grow 6 days at a subtoxic concentration of ATRA and then treated with different concentration of CDDP and 5-FU for 24 h. The cytotoxicity was examined by cell proliferation MTT assay. Additionally, AO/EB staining was used for detection of apoptotic cells. In order to determine whether the growth inhibition was also associated with changes in cell cycle distribution, cell cycle analysis was performed using flow cytometry.Low concentration of ATRA (1 μM, 6days) followed by 5-FU and CDDP was found to be more effective than either drugs alone, thus resulting in synergistic cytotoxicity in Kyse-30 and AGSCD44^± cells. Furthermore, there was an indication that the combination of ATRA with 5FU and CDDP caused an increase in cell cycle arrest in G2/M and G0/G1.We conclude that low concentration of ATRA enhances the cytotoxicity of CDDP and 5FU by facilitating apoptosis and cell cycle arrest in gastrointestinal CSCs and provide a rational basis for the design of novel, well-tolerated CDDP- and 5FU-based chemotherapy in human gastrointestinal carcinoma.     

针对MAGE-1基因RNAi在恶性胶质瘤SHG-44细胞中的作用研究

目的构建抑制黑色素瘤抗原-1(MAGE-1)的siRNA表达载体,鉴定其在人恶性胶质瘤细胞系SHG-44细胞中对MAGE-1基因表达的干涉作用。方法化学合成2对编码短发夹RNA序列的靶向MAGE-1基因寡核苷酸链,克隆到经BglⅡ、HindⅢ双酶切的pSUPER载体上,重组构建RNA干涉(RNAi)质粒载体。利用逆转录多聚酶链反应(RT-PCR)、流式细胞术和荧光显微镜,检测经稳定转染后SHG-44细胞中MAGE-1的表达,以了解siRNA的干扰效果。结果重组构建的pSUPER-MAGE-1载体经双酶切电泳及插入基因片段序列分析,表明寡核苷酸链成功插入到预计位点,并且序列与预期完全一致。稳定转染后G418筛选出的SHG-44多克隆细胞MAGE-1的表达经RT-PCR、流式细胞术和荧光显微镜检测,2对siRNA均有较明显的干涉作用。结论成功构建了针对MAGE-1基因的siRNA表达载体,抑制SHG-44细胞中的MAGE-1分子的表达。     

CD44v7/8基因表达与宫颈癌关系的探讨

目的探讨宫颈癌组织中CD44v7/8基因的异常表达及临床意义。方法1986～2000年沈阳医学院附属中心医院采用免疫组化的方法随机测定59例宫颈癌、18例尖锐湿疣和18例慢性宫颈炎组织中CD44v7/8基因的表达情况。结果宫颈癌、尖锐湿疣及慢性宫颈炎3种疾病中CD44v7/8表达阳性率分别为76.27%、11.11%和11.11%,宫颈癌组的CD44v7/8表达分别高于慢性宫颈炎组及尖锐湿疣组,且差异非常显著(P<0.01)。慢性宫颈炎组及尖锐湿疣组比较差异无显著性(P>0.05)。结论CD44v7/8蛋白的表达增强与宫颈癌的发生、发展及和局部浸润转移有关。     

CD44分子与乳腺癌

CD44分子属于黏附分子家族中的一员，其CD44的异常表达与乳腺癌的发生、发展和转移密切相关。近年来研究表明乳腺癌干细胞表面高度表达CD44分子，针对CD44分子与乳腺癌干细胞关系的研究，将为乳腺癌的临床治疗提供新的靶点。     

三氧化二砷对CO2气腹下小鼠肿瘤腹壁戳口种植影响的实验研究

目的 研究腹腔内注射三氧化二砷(arsenic trioxide,As2O3)对小鼠CO2气腹下肝癌H22转移的影响. 方法 昆明鼠40只(清洁级),中腹部穿刺置入1 mm套管针,自套管针注入1×106肿瘤细胞后,建立CO2气腹,压力8 mm Hg,时间30 min.术后随机分4组,每组10只,分别腹腔内注入生理盐水,1 ml;As2O3(2 mg/kg),1 ml;As2O3(4 mg/kg),1 ml;As2O3(4mg/kg)+肝素(10 U/ml),共1 ml.气腹后第3、7天测量肿瘤黏附因子(CD44)、血管内皮生长因子(vascular endothelial growth factor,VEGF)的变化;比较各组生存状态、腹围、体重变化及转移瘤直径.结果 气腹后第3、7天,与对照组相比,各As2O3组CD44、VEGF表达均明显降低(P＜0.05).2个高剂量组的气腹后第3天VEGF、第7天CD44比低剂量组降低明显(P＜0.05).4组戳口种植率分别为9/10、8/10、7/10、6/10,差异无显著性(x2=2.667,P=0.446). 结论 As2O3对CO2气腹腹腔镜肿瘤生长转移有抑制作用.     

BcL-2、Bax、CD44V6在乳腺癌表达及预后的研究

目的探讨影响乳腺癌患者预后的因素,协助临床制定手术方式及术后治疗方案。方法选择80例各型乳腺癌患者的标本,应用免疫组化SP方法,检测乳腺癌组织中癌基因BcL-2、Bax及黏附因子CD44V6的表达情况。结果乳腺癌中BcL-2、Bax及CD44V6的表达情况与肿瘤组织学分级、瘤体直径。有无转移及患者术后存活时间均有显著性相关(P<0.05或0.01)。结论BcL-2高表达、Bax低表达、CD44V6低表达的患者组织学分级好,肿瘤体积小,淋巴结转移少,术后存活时间长,这部分患者可做肿物扩大切除而保留乳房或即使已做乳房切除,术后也可减少放疗及化疗剂量。     

HLA-B*27 subtyping by PCR-RFLP in Spanish patients with ankylosing spondyiitis

Abstract: We describe the use of restriction analysis on PCR-amplified DNA for detecting all B*27 subtypes except B*2710 and B*2711 (i.e. from B*2701 to B*2709). After detecting B*27 by Sty I, double digestions consisting of Sty I plus another informative enzyme led to subtype assignment. We used mismatched primers to create restriction sites when necessary. The method avoids group-specific amplifications and other laborious optimization procedures. It was successfully tested on a panel of well characterized cell lines covering different B*27 subtypes. Then, we studied a group of 57 ankylosing spondyiitis patients and 746 controls from the south of Spain. B*27 showed a very strong association with the disease (OR=211.27, P=\0˜7). B*2702 and B*2705 distribution in controls (20% and 77.1%, respectively) differed from previously reported data in the Spanish population. We unexpectedly found the B*2707 allele in our population (one control).