PTEN、nm23、bcl-2、P53与胃癌细胞转移的临床研究
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摘要
研究目的:研究胃癌组织中P53、nm23、bcl-2、PTEN基因的表达与临床病理的关系,探讨p53,nm23,bcl-2,PTEN基因与胃癌分期,转移的关系。
方法:应用免疫组织化学LSAB法检测85例胃癌病人nm23,bcl-2、P53基因表达情况,应用PCR-SSCP法检测45例病人是否存在p53基因的第七外显子突变和P7EN基因突变,对3例存在PTEN基因突变的病人分别行第五和第八外显子DNA测序。
结果:85例病人,P53阳性率为41.18%,早期胃癌病人占15.38%,进展期胃癌P53阳性率为45.83%,两者显著差异(X~2=4.097,P<0.05),有淋巴结转移和无淋巴结转移者高度差异性,(X~2=11.016,p<0.001),有淋巴结转移者P53表达强度高于无淋巴结转移者(u=3.055 P<0.01),进展期胃癌P53表达强度高于早期胃癌(u=1.972 p<0.005);P53基因第七外显子突变发生率低分化腺癌显著高于高、中分化腺癌(p=0.0085),有淋巴结转移者高于未发生淋巴结转移者(p=0.043),与浸润深度无关(p>0.05)。bcl-2整体阳性率为52.94%,早期胃癌显著高于进展期胃癌(X~2=6.1803 p<0.05),有淋巴结转移者同无淋巴结转移者之间无差异(X~2=0.4561 p>0.05),bcl-2表达与胃癌分化程度之间无差异(X~2=0.3860p>0.05),不同细胞类型间无差异(X~2=1.5921 p>0.05);nm23-H1蛋白总体阳性率为56.47%,早期胃癌高于进展期胃癌(X~2=7.6737 p<0.01),无淋巴结转移者显著高于有淋巴结转移者(X~2=7.6937 p<0.01)。PTEN基因突变率为7.14%(3/42),高分化癌变组织未见突变,不同临床分期间无差异(X~2=0.1216 p=0.7273),胃
胃癌转移P53
n们n23
bCI一2
PTEN
PCR一SSCP
Objective: To study the relationship among the expression of PTEN、 nm23、 bcl-2 and p53 and clinicopathology in the patients with gastric cancer. Method 85 cases of different differentiated gastric cancer, 11 cases of well-differentiated gastric cancer, 29 cases of mid-differentiated gastric cancer and 45 cases of poor-differentiated gastric cancer, were stained by immunohistochemical LSAB method.45 cases were studied by PCR-SSCP to detect the mutational differentiation genic mutation in p53 (exon7) and PTEN (exon5、 exon8) . 3 cases with PTEN mutation were further studied with DNA sequence assay. Results The positive rates of p53 protein expression was 41.18% , there were significant deviation between early gastric cancer (15.38%) and progressive gastric cancer (45.83%) (X2=4.097 P<0.05), the difference exsit significantly between patients with lymphatic metastasis and non-lymphatic metastasis (x2=11.036, p<0.001) ; The positive density of p53 was higher in patients with lymphatic metastasis and in patients of progressive gastric cancer significantly than those of patients with non-lymphotic metastasis (u=3.055 p<0.01) and early gastric cancer (u=1.972 p<0.005);P53 mutation was more frequently in poor-differentiated gastric cancer than patients with well- differentiated gastric cancer (p=0.0085) and with lymphatic metastasis(p=0.043),and had no relation
with the deepth of infiltration. The positive rates of bcl-2 protein expression was 52.94% ,that of patients with progressive gastric cancer was significanty higher than that with early gastric cancer(x2=6.1803 p<0.05),and had no relation with lymphatic metastasis (x2=0.4561 p>0.05), differentiated degree(x2=0.386 p>0.05) and cellular types(x2= 1.5921 p>0.05).The positive rate of nm23 protein expression was 56.47%,and it was significantly higher in progressive gastric cancer and patients with non lymphatic metastasis than that in early gastric cancer(x2=7.6737 p<0.01) and those with lymphatic metastasis(x2=7.6937 p<0.01).The mutatual rate of PTEN was 7.14%(3/42),there were no significant difference in groups of different clinical stages(x2=0.1216 p=7273) and different differentiated degree(x2=0.7602 p=0.3833). PTEN mutation DNA sequence assay found A basic deletion at 25point in exon5,G mutae to C C mutate to A at 937 and 992 in exon8 differently .Patients with p53(+) and nm23(-) were significantly apt to occur lymphatic metastasis than those with p53(-) and nm23(+)(x2=5.036 p<0.05),no difference were found in other groups (p>0.05).Conclusion:1.p53 nm23 bcl-2 protein expression had synergistic effect to the gastric metastasis;2.the p53 genic mutation assay may be helpful to identify the patients with high malignant gastric cancer;3.PTEN mutations were apt occur in poor-differentiated gastric cancer . 4.PTEN p53 nm23 and bcl-2 co-assay was valuable to predict the prognosis of patients.
方法:应用免疫组织化学LSAB法检测85例胃癌病人nm23,bcl-2、P53基因表达情况,应用PCR-SSCP法检测45例病人是否存在p53基因的第七外显子突变和P7EN基因突变,对3例存在PTEN基因突变的病人分别行第五和第八外显子DNA测序。
结果:85例病人,P53阳性率为41.18%,早期胃癌病人占15.38%,进展期胃癌P53阳性率为45.83%,两者显著差异(X~2=4.097,P<0.05),有淋巴结转移和无淋巴结转移者高度差异性,(X~2=11.016,p<0.001),有淋巴结转移者P53表达强度高于无淋巴结转移者(u=3.055 P<0.01),进展期胃癌P53表达强度高于早期胃癌(u=1.972 p<0.005);P53基因第七外显子突变发生率低分化腺癌显著高于高、中分化腺癌(p=0.0085),有淋巴结转移者高于未发生淋巴结转移者(p=0.043),与浸润深度无关(p>0.05)。bcl-2整体阳性率为52.94%,早期胃癌显著高于进展期胃癌(X~2=6.1803 p<0.05),有淋巴结转移者同无淋巴结转移者之间无差异(X~2=0.4561 p>0.05),bcl-2表达与胃癌分化程度之间无差异(X~2=0.3860p>0.05),不同细胞类型间无差异(X~2=1.5921 p>0.05);nm23-H1蛋白总体阳性率为56.47%,早期胃癌高于进展期胃癌(X~2=7.6737 p<0.01),无淋巴结转移者显著高于有淋巴结转移者(X~2=7.6937 p<0.01)。PTEN基因突变率为7.14%(3/42),高分化癌变组织未见突变,不同临床分期间无差异(X~2=0.1216 p=0.7273),胃
胃癌转移P53
n们n23
bCI一2
PTEN
PCR一SSCP
Objective: To study the relationship among the expression of PTEN、 nm23、 bcl-2 and p53 and clinicopathology in the patients with gastric cancer. Method 85 cases of different differentiated gastric cancer, 11 cases of well-differentiated gastric cancer, 29 cases of mid-differentiated gastric cancer and 45 cases of poor-differentiated gastric cancer, were stained by immunohistochemical LSAB method.45 cases were studied by PCR-SSCP to detect the mutational differentiation genic mutation in p53 (exon7) and PTEN (exon5、 exon8) . 3 cases with PTEN mutation were further studied with DNA sequence assay. Results The positive rates of p53 protein expression was 41.18% , there were significant deviation between early gastric cancer (15.38%) and progressive gastric cancer (45.83%) (X2=4.097 P<0.05), the difference exsit significantly between patients with lymphatic metastasis and non-lymphatic metastasis (x2=11.036, p<0.001) ; The positive density of p53 was higher in patients with lymphatic metastasis and in patients of progressive gastric cancer significantly than those of patients with non-lymphotic metastasis (u=3.055 p<0.01) and early gastric cancer (u=1.972 p<0.005);P53 mutation was more frequently in poor-differentiated gastric cancer than patients with well- differentiated gastric cancer (p=0.0085) and with lymphatic metastasis(p=0.043),and had no relation
with the deepth of infiltration. The positive rates of bcl-2 protein expression was 52.94% ,that of patients with progressive gastric cancer was significanty higher than that with early gastric cancer(x2=6.1803 p<0.05),and had no relation with lymphatic metastasis (x2=0.4561 p>0.05), differentiated degree(x2=0.386 p>0.05) and cellular types(x2= 1.5921 p>0.05).The positive rate of nm23 protein expression was 56.47%,and it was significantly higher in progressive gastric cancer and patients with non lymphatic metastasis than that in early gastric cancer(x2=7.6737 p<0.01) and those with lymphatic metastasis(x2=7.6937 p<0.01).The mutatual rate of PTEN was 7.14%(3/42),there were no significant difference in groups of different clinical stages(x2=0.1216 p=7273) and different differentiated degree(x2=0.7602 p=0.3833). PTEN mutation DNA sequence assay found A basic deletion at 25point in exon5,G mutae to C C mutate to A at 937 and 992 in exon8 differently .Patients with p53(+) and nm23(-) were significantly apt to occur lymphatic metastasis than those with p53(-) and nm23(+)(x2=5.036 p<0.05),no difference were found in other groups (p>0.05).Conclusion:1.p53 nm23 bcl-2 protein expression had synergistic effect to the gastric metastasis;2.the p53 genic mutation assay may be helpful to identify the patients with high malignant gastric cancer;3.PTEN mutations were apt occur in poor-differentiated gastric cancer . 4.PTEN p53 nm23 and bcl-2 co-assay was valuable to predict the prognosis of patients.
引文
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2. Oue N, Hamai Y, Mitani Y, et al. Gene expression profile of gastric carcinoma: identification of genes and tags potentially involved in invasion, metastasis, and carcinogenesis by serial analysis of gene expression. Cancer Res. 2004 Apr 1;64(7):2397-405.
3.冯建芳,章静波.程序性细胞死亡及细胞凋亡.生理科学进展,1995,26(4):373-378.
4. Quinlan DC, Davidson AG, Summers CL, et al. Accumulation of P53 protein correlated with a poor prognosis in human lung cancer . Cancer Res, 1992, 52: 4828-4833.
5. Barnes DM, Dublin EA, Fisher CJ, et al. Immunohistochemical detection of P53 protein in mammary carcinoma: an important new independent indicator of prognosis? Hum Pathol, 1993, 24:469-476.
6. Brito MJ, Williams GT, Thompson H. Expression of p53 in early (T1)gastric carcinoma and precancerous adjacent mucosa. Gut, 1994,35:1697-1700.
7.吴黎华,李日镛,金昌范,等.突变型p53和c-neu癌基因产物在胃癌中的表达.临床与实验病理学杂志,1997;13(1):30-34.
8. Waldman V, Kinzler KW, Vogelstein B. P21 is necessary for the p53-mediated G1 arrest in human cancer cells. Cancer Res, 1995,55:5187-5190.
9. Fritsche M, Haessler C,Brandner C. Induction of nuclear accumulation of the tumor-suppressor protein p53 by DNA-damaging agents. Oncogene, 1993, 8: 307-310.
10. Orita M, Suzuki Y, Sekiya T, et al. Rapid and sensitive detection of point mutation and DNA polymorphisms using the polymerase chain reaction. Proc Natl Acad Sci USA, 1989, 87:874-878.
11. Orita M, Iwahana H, Kanazawa H, et al. Detection of polymorphisms of human DNA by gel electrophoresis as single-strand conformation polymorphisms. Proc Natl Acad Sci USA, 1989, 86:2766-2769.
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