Differential analysis of lymph node metastasis in histological mixed-type early gastric carcinoma in the mucosa and submucosa

AIM To investigate the relationship between histological mixed-type of early gastric cancer(EGC) in the mucosa and submucosa and lymph node metastasis(LNM).METHODS This study included 298 patients who underwent gastrectomy for EGC between 2005 and 2012. Enrolled lesions were divided into groups of pure differentiated(pure D), pure undifferentiated(pure U), and mixed-type according to the proportion of the differentiated and undifferentiated components observed under a microscope. We reviewed the clinicopathological features, including age, sex, location, size, gross type, lymphovascular invasion, ulceration, and LNM, among the three groups. furthermore, we evaluated the predictors of LNM in the mucosa-confined EGC.RESULTS Of the 298 patients, 165(55.4%) had mucosa-confined EGC and 133(44.6%) had submucosa-invasive EGC. Only 13(7.9%) cases of mucosa-confined EGC and 30(22.6%) cases of submucosa-invasive EGC were observed to have LNM. The submucosal invasion(OR = 4.58, 95%CI: 1.23-16.97, P = 0.023), pure U type(OR = 4.97, 95%CI: 1.21-20.39, P = 0.026), and mixedtype(OR = 5.84, 95%CI: 1.05-32.61, P = 0.044) were independent risk factors for LNM in EGC. The rate of LNM in mucosa-confined EGC was higher in the mixedtype group(P = 0.012) and pure U group(P = 0.010) than in the pure D group, but no significant difference was found between the mixed-type group and pure U group(P = 0.739). Similarly, the rate of LNM in the submucosa-invasive EGC was higher in the mixedtype(P = 0.012) and pure U group(P = 0.009) than in the pure D group but was not significantly different between the mixed-type and pure U group(P = 0.375). Multivariate logistic analysis showed that only female sex(OR = 5.83, 95%CI: 1.64-20.70, P = 0.028) and presence of lymphovascular invasion(OR = 13.18, 95%CI: 1.39-125.30, P = 0.020) were independent risk factors for LNM in mucosa-confined EGC, while histological type was not an independent risk factor for LNM in mucosa-confined EGC(P = 0.106).CONCLUSION for mucosal EGC, histological mixed-type is not an independent risk factor for LNM and could be managed in the same way as the undifferentiated type.

IV型胶原纤维及其酶在胃癌浸润转移中的作用

采用ＩＶ型胶原纤维单抗和ＩＶ型胶原酶多抗检测７６例胃癌。发现ＩＶ型胶原纤维染色所示基底膜明显缺损，在分化较差者及浸润型胃癌中（＞７５％）明显高于分化较好者及膨胀型胃癌（Ｐ＜０．００５）。ＩＶ型胶原酶明显增加在分化较差者，浸润型及有淋巴结转移者（８０％左右）显著高于分化较好者，膨胀型及无淋巴结转移者（Ｐ＜０．０５）。说明分化程度低的胃癌产生更多ＩＶ型胶原酶，严重破坏基底膜，促进浸润和淋巴结转移。其可作为恶性程度指标。

IV型胶原酶在不同转移潜能的前列腺癌细胞系中的表达及活性分析

目的:探讨前列腺癌细胞转移潜能与IV型胶原酶基质金属蛋白酶2(MMP-2)和基质金属蛋白酶9(MMP-9)的关系。方法:应用免疫组织化学SABC法,研究IV型胶原酶在不同转移潜能的前列腺癌细胞株LNCaP、C4-2B中表达变化。同时采用明胶酶谱法对这2种前列腺癌细胞系分泌的IV型胶原酶活性进行分析。结果:①免疫组织化学染色显示,LNCaP、C4-2B细胞中均有MMP-2、MMP-9表达,主要表达于细胞质中,C4-2B细胞表达明显强于LNCaP(P<0.01)。②人前列腺癌细胞LNCaP的条件培养液中仅能检测到极低的MMP-2和MMP-9活性,酶谱条带的积分A值分别为0.89±0.02和0.86±0.01,主要以酶原形式存在,而C4-2B细胞则表现较强的MMP-2、MMP-9活性,积分A值分别为96.32±4.36和33.89±1.84,其分泌的MMP-2则以活性形式为主,其分泌量显著高于LNCaP细胞(P<0.01)。结论:前列腺癌细胞中IV型胶原酶的表达和分泌,有助于癌细胞的侵袭和转移;前列腺癌细胞的侵袭转移潜能与其产生IV型胶原酶的能力密切相关。

Intracellular expression of a single-chain antibody directed against type IV collagenase inhibits the growth of lung cancer xenografts in nude mice

It was documented that type IV collagenase with two subtypes of 72 ku/MMP-2 and 92 ku/MMP-9 plays an important role in tumor invasion and metastasis. The endoplasmic reticulum (ER)- retained, single chain Fv antibody fragment (scFv) was used to inhibit the function of type IV collagenase. For expression in mammalian cells, the assembled scFv M97 gene with ER retention signal encoding 6 additional amino acids (SEKDEL) was reamplified by PCR. The amplified fragments were cloned into the pcDNA3.1 vector. The resulting plasmid was sequenced and then introduced into PG cells, a highly metastatic human lung cancer cell line, by lipofectAMINE method. The result of intrabody gene therapy showed that type IV collegenase expression was down regulated significantly as measured by ELISA. The biological behavior of PG cell, such as the ability of in vitro invasion through Matrigel, colony formation on soft agar, was also inhibited by scFv M97 transfection. Animal experiments in a xenograft model of human lung

肝癌患者血清Ⅳ型胶原降解酶活性与转移的关系

为了解基质降解在原发性肝癌转移中的作用，检测了６５例原发性肝癌患者的血清Ⅳ型胶原降解酶活性。结果表明：原发性肝癌患者血清Ⅳ型胶原降解酶活性明显高于肝硬化患者及正常对照；发生肝内、门脉转移者，酶活性高于无转移者；出现远处转移者酶活性明显高于肝内、门脉转移者。对６例患者的动态观察表明，随着肝内、门脉转移，远处转移的发生，酶活性显著升高。肝癌患者血清Ⅳ型胶原降解酶活性与ＡＦＰ、肿瘤大小无相关性。

胃癌组织中CD44v6蛋白表达和血清sCD44v6水平变化的研究

目的研究组织中CD44v6的表达以及血清sCD44v6水平变化与胃癌生物学行为和Lauren分型的关系,为进一步探讨两者与胃癌浸润转移关系奠定基础。方法采用免疫组化SP法观察CD44v6在42例胃癌组织中的表达情况,以及应用ELISA法检测53例胃癌患者血清sCD44v6水平变化。结果CD44v6在30例正常胃粘膜组织中均不表达,在胃癌组织中表达61.9%(2642),血清sCD44v6在胃癌组(n=53)水平为(565.69±165.89)ng ml明显高于正常对照组(n=42)及胃溃疡组(n=29),无论CD44v6的表达还是sCD44v6水平均随胃癌浸润扩展、TNM分期上升及转移的发生而增高。弥漫型胃癌CD44v6表达80%(1620)明显高于肠型胃癌者45.5%(1022),并且随着浸润进展及转移程度而增加(P<0.05),同时弥漫型胃癌患者血清sCD44v6水平[(629.61±118.10)ng ml,n=22]明显高于肠型胃癌者[(520.33±101.17)ng ml,n=31],并随着远处转移的发生而增高(P<0.05)。结论组织中CD44v6表达和血清sCD44v6水平变化在胃癌,尤其是弥漫型胃癌的浸润转移的预测与监测具有临床检测意义。

混合型组织学类型胃癌组织中CD44v6表达与侵袭和转移的关系

目的 研究CD4 4v6在不同组织学类型的胃癌和混合型组织学类型胃癌中表达与侵袭和转移的关系。方法 4 0例经外科手术切除的胃癌石蜡包埋组织 ,选用CD4 4v6单克隆抗体 ,采用sABC法 ,进行免疫组织化学检测。结果 中分化腺癌常伴有粘液腺癌 ;低分化腺癌多伴有印戒细胞癌。CD4 4v6在中分化腺癌伴粘液腺癌组织中的表达率为 33 3% ;而在低分化腺癌伴印戒细胞癌组织中的表达率为 6 6 7%。结论 低分化腺癌伴有印戒细胞癌中CD4 4v6的高表达率可能说明在侵袭和转移方面高于中分化腺癌伴有的粘液腺癌。

TIMP-2基因转染裸鼠胃癌中Ⅳ型胶原酶及Ⅳ型胶原改变的研究

用金属蛋白酶抑制剂——TIMP-2基因体外转染的胃癌SGC-7901细胞系建立裸鼠肾包膜下种植的侵袭模型,并应用免疫组化对侵袭部位的组织Ⅳ型胶原酶前体——MT-MMP的表达及基底膜的改变作了研究.结果显示,正常肾脏组织Ⅳ型胶原酶反应阳性率为39.64%,在种植肿瘤中阳性率为69.14%,两者差异显著(P<0.01),Ⅳ型胶原染色显示,肾小管及肾小球基底膜完整,而肿瘤中基底膜出现断裂或缺失,在肿瘤浸润部见Ⅳ型胶原酶表达异常丰富,而Ⅳ型胶原缺如.表明Ⅳ型胶原酶对基底膜的破坏是胃癌浸润转移的重要机制.

蛋白激酶C对大肠癌细胞转移特性调控的实验研究

目的:探讨蛋白激酶C(protein kinase C.PKC)对大肠癌细胞转移特性的调控作用。方法:采用羊膜侵袭培养系统和明胶酶谱分析的方法,研究了PKC激活剂PMA对人结肠癌细胞株HT-29体外侵袭作用的影响及PKC抑制剂Staurosporine(sP)对PMA的拮抗作用,以及研究了这种体外的侵袭作用与细胞分泌Ⅳ型胶原酶,包括72kDⅣ型胶原酶(MMP-2)和92kDⅣ型胶原酶(MMP-9)的关系。结果:PMA可显著增强HT-29细胞的侵袭性,与对照组相比,具有显著性差异(P<0.01).而SP则可拮抗PMA的这种诱导作用。PMA还可增加HT-29细胞分泌MMP-2和MMP-9,而SP则可拮抗PMA的这种诱导作用,抑制MMP-2和MMP-9的分泌。结论:蛋白激酶C可能是调控肿瘤细胞侵袭、转移的重要因素。PKC的激活可诱导肿瘤细胞侵袭性增强和分泌MMP-2和MMP-9增加,SP可能通过抑制PMA对PKC的激活,进而抑制了肿瘤细胞的侵袭和分泌MMP-2和MMP-9。MMP-2和MMP-9的分沁与肿瘤细胞侵袭性有密切关系。

Ⅳ型胶原酶在胃癌浸润转移中的作用

应用免疫组化技术研究４６例胃癌手术切除新鲜标本及其引流区域淋巴结中Ⅳ型胶原酶的表达，以探索其在胃癌浸润转移中的作用。结果发现：Ⅳ型胶原酶存在于正常胃小凹上皮和胃癌中，但在５７％（２６／４６）胃癌中的表达增强（Ｐ＜０．０１），而且在有些胃癌组织浸润前缘其表达增强，在胃癌原发灶中Ⅳ型胶原酶的表达和淋巴结转移灶中的表达一致。另外，在有淋巴结转移的胃癌原发灶中，Ⅳ型胶原酶的表达较无淋巴结转移的胃癌原发灶中的表达明显增强（Ｐ＜０．０１）．实验表明，Ⅳ型胶原酶可能与胃癌浸润转移密切相关，它可以作为判断胃癌浸润转移恶性表型的有用生物学标志。