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中国图书资料分类法分类号R735.2
The role of cyclin E expression in the intestinal type gastric cancer
LI Jia-Qing, CAI Wen-Yan, ZHANG Zai-Xing
(Department of Surgery, the First Hospital, Beijing Medical University, Beijing100034)
MeSHcyclins/biosynStomach neoplasms/chemcyclin E☆
ABSTRACTObjective: To investigate the correlation of cyclin E expression with the clinicopathologic features and prognosis of intestinal type gastric cancer.Methods: The expression of cyclin E protein in 59 patients with intestinal type gastric cancer was studied by means of immunohistochemistry.Results: Normal gastric epithelial cells were negative, while 30(30/59) carcinomatous tissues were immunostained. Abnormal expression of cyclin E was significantly related to invasion depth of intestinal type gastric cancer. The patients with strongly positive expression of cyclin E tended to have a lower postoperative 5 year survival rate than other groups.Conclusion: Abnormal expression of cyclin E might take part in and promote the progression of intestinal type gastric cancer, and it might be used as a prognostic factor.
（J Beijing Med Univ, 1999，31：71-73）
人类细胞中，cyclin E、cyclin A、cyclin D是G1期细胞周期蛋白。cyclin E、细胞周期蛋白依赖激酶2(cyclin-dependent kinase 2，CDK2)共同在G1晚期调节G1期到S期的过渡。cyclin E结合并激活CDK2可直接促进G1/S期的转变。异常变化的cyclin E将使G1/S期过渡失调，有可能导致细胞的无限制增殖，而参与多种肿瘤的发生和发展［1］。文献报道cyclin E的异常表达能促进结、直肠癌的发生和进展［2］，并与乳腺癌的发生相关［3］。本文探讨cyclin E在肠型胃癌中的表达及其与肠型胃癌预后之间的关系。
1材料与方法
取1987～1991年连续手术切除的肠型胃癌标本59例。其中男47例，女12例；年龄最大88岁，最小45岁。随访至1996年12月30日。标本经100 g.L－1福尔马林固定，石蜡包埋。每例标本都选择包括肿瘤中心和肿瘤边缘的石蜡块1～2个，制成连续切片后，进行免疫组化和HE染色。肠型胃癌的组织学类型包括乳头状腺癌、管状腺癌和粘液腺癌。
3 μm厚的切片按ABC法进行免疫组化染色［2］，药盒为Histofine (Nichirei公司，日本)产品。组织切片经二甲苯脱蜡、水化后浸入0.088mol.L－1的H2O2中30 min，以阻断内源性氧化酶的活性。为充分暴露抗原，切片在0.01 mol.L－1、pH 6.5的柠檬酸缓冲液内加热到100℃，持续30min。经抗cyclin E单克隆抗体(Santa Cruz公司，体积比1∶600稀释)4℃孵育12 h后，室温下切片分别再与兔抗鼠血清反应20 min，PAP(peroxidase-anti-peroxidase)复合物反应15 min。随后H2O2-DAB显色2 min。经苏木精复染1 min，脱水、透明、封片。
每张切片选择5个不同的代表性区域（即切片的中心及上、下、左、右四边）分别进行细胞计数。每区计数200、5区共1 000个细胞，算出染色细胞所占比例。cyclin E的染色结果分为3级：阴性(－)，染色细胞数＜5％；阳性(+)，染色细胞数5％～50％；强阳性(++)，染色细胞数＞50％｛1］。
统计学处理：cyclin E表达与预后的关系使用Kaplan-Meier分析中的Longrank法，与浸润深度、性别等的关系采用χ2检验，与年龄等的关系使用双侧t检验。
2结果
2.1正常和癌组织中cyclin E的表达
正常胃粘膜上皮细胞不被抗cyclin E单克隆抗体染色。各种血管内皮细胞浆都呈棕色反应，淋巴细胞浆轻度黄染，而两种细胞胞核均不染色（图1）。肠型胃癌组织中，棕色的免疫反应物位于细胞核内，且染色细胞分布不均匀（图2）。肠型胃癌总的染色阳性共有30例(30/50)，其中(+)有18例(18/59)，(++)有12例(12/59)，表1。
图1正常胃粘膜cyclin E表达：腺上皮细胞不被染色，
部分淋巴细胞浆轻度黄染免疫组化染色×200
Figure 1Expression of cyclin E in normal gastric mucosa. None
of glandular epithelial cell was stained. The cytoplasm of some
lymphocytes was slightly stained brown
Immunohistochemical staining×200
图2肠型胃癌组织cyclin E表达：棕色免疫反应物位于
细胞核内，染色细胞所占比例≥5%免疫组化染色×200
Figure 2Expression of cyclin E in gastric cancerous tissue. The brown
immunoreactivities were localized in the nuclei of stained cells.
Stained cells were more than 5% of counted cells
Immunohistochemical staining×200
2.2cyclin E蛋白表达与临床病理特征的关系
cyclin E蛋白表达与患者年龄、肿瘤大小、位置、病理肉眼类型、组织学类型、间质量、淋巴结转移和腹膜转移无关。但与性别相关，男性患者表达增高；其表达强度与肿瘤的浸润深度也相关，浸润深者表达增高；另外，异常表达还与肠型胃癌的远处转移有关，强阳性表达患者的远处转移率显著增高，表1。
表1肠型胃癌中cyclin E的表达与临床病理特征的关系
Table 1The correlation of cyclin E expression
with clinicopathologic features in intestinal type gastric cancer
Clinicopathologicfeatures n cyclin E expression － ＋ ＋＋ TPR／% Sum 59 29 18 12 51 Sex Male 47 20 17 10 57 Female 12 9 1 2 25 Invasion depth Mucosa 12 9 3 0 25 Submucosa 14 4 8 2 71 Muscularis 9 3 3 3 67 Other* 24 13 4 7 46 Remote Negative 56 29 18 9 48 metastasis Positive 3 0 0 3 100
TPR，total positive rate; *,included subserosa, serosa and metastasis; sex,male vs female, P＜0.05; invasion depth, mucosa vs submucosa, P＜0.05; mucosa vs muscularis, P=0.059; mucosa vs other, P＞0.05; submucosa vs muscularis, P＞0.05; submucosa vs other, P＜0.05; muscularis vs other, P＞0.05; remote metastasis, negative vs positive, P＜0.01.
2.3cyclin E蛋白表达与预后的关系
在全部肠型胃癌患者中强阳性表达者术后5年生存率显著降低。虽差异无统计学意义，但根治性切除患者中强阳性表达组的5年生存率仍然较低(表2)。
表2肠型胃癌中cyclin E表达与预后的关系
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