目的:检测大肠癌原发灶与淋巴结(lymph node,LN)转移灶中磷脂酰肌醇3-激酶(phosphatidylinositol 3-kinase,PI3K)p110α、PI3Kp110β与B细胞淋巴瘤基因-2(B c e l l lymphoma 2,Bcl-2)、CyclinD1的表达情况,探讨在大肠癌发生及转移中的...目的:检测大肠癌原发灶与淋巴结(lymph node,LN)转移灶中磷脂酰肌醇3-激酶(phosphatidylinositol 3-kinase,PI3K)p110α、PI3Kp110β与B细胞淋巴瘤基因-2(B c e l l lymphoma 2,Bcl-2)、CyclinD1的表达情况,探讨在大肠癌发生及转移中的相关性及其与临床病理因素及预后的关系.方法:采用免疫组织化学方法在30例正常大肠黏膜组织,52例未发生LN转移的大肠癌组织,50例发生LN转移的大肠癌原发灶及其转移灶中检测PI3Kp110α、PI3Kp110β、Bcl-2和CyclinD1的表达情况及其差异,分析PI3Kp110α、PI3Kp110β与Bcl-2和CyclinD1之间的相关性以及与临床病理因素及其预后的关系.结果:(1)PI3Kp110α与Bcl-2在无LN转移组、有LN转移组的原发灶和其相应LN转移灶中的表达均高于正常肠黏膜组(P<0.05);PI3Kp110β与CyclinD1在无LN转移组、有LN转移组的原发灶和其相应LN转移灶中的表达均高于正常肠黏膜组,且在有LN转移的原发灶中的表达均高于其相应LN转移灶和无LN转移组肠癌中的表达(P<0.05);(2)PI3Kp110α与Bcl-2在4组中均呈正相关(P<0.05);PI3Kp110α与CyclinD1在正常肠黏膜组,无LN转移组和有LN转移组中均呈正相关(P<0.05);P I3Kp110β与Bcl-2和CyclinD1在4组中均呈正相关(P<0.05);(3)PI3Kp110α、PI3Kp110β和CyclinD1蛋白的表达与肿瘤分化程度及LN转移相关,Bcl-2的表达与肿瘤分化程度相关;(4)Kaplan-Meier分析显示:PI3Kp110α、PI3Kp110β、Bcl-2和CyclinD1为影响大肠癌预后的因素之一;Cox比例风险模型分析显示:PI3Kp110α、PI3Kp110β是影响大肠癌患者预后的独立因素.结论:(1)PI3Kp110α、PI3Kp110β与Bcl-2、CyclinD1在大肠癌原发灶及转移灶中的表达均高于正常黏膜,在肿瘤的发生发展中发挥重要作用;(2)大肠癌LN转移灶中,PI3Kp110α与PI3Kp110β分别通过影响Bcl-2及CyclinD1对转移灶中的肿瘤生长起促进作用;(3)PI3Kp110α、PI3Kp110β、Bcl-2和CyclinD1与大肠癌的分化程度有关,且PI3Kp110α、PI3Kp110β和CyclinD1与大肠癌的转移密切相关;(4)PI3Kp110α和PI3Kp110β是影响大肠癌预后的独立危险因素.展开更多
This study examined the synergetic effect of class ⅠA Phosphoinositide 3-kinases catalytic subunit p110β knockdown in conjunction with oxaliplatin treatment on colon cancer cells. Down-regulation of p110β by siRNA ...This study examined the synergetic effect of class ⅠA Phosphoinositide 3-kinases catalytic subunit p110β knockdown in conjunction with oxaliplatin treatment on colon cancer cells. Down-regulation of p110β by siRNA interference and oxaliplatin treatment were applied in colon cancer cell lines HT29, SW620 and HCT116. MTT assay was used to measure the inhibitory effect of p110β knockdown on the proliferation of colon cancer cell lines. SubG1 assay and Annexin-Ⅴ FITC/PI double-labeling cytometry were applied to detect cell apoptosis. And cell cycle was evaluated by using PI staining and flow cytometry. The expression of caspase 3, cleaved PARP, p-Akt, T-Akt and p110β was determined by western blotting. The results suggested that down-regulation of p110β expression by siRNA obviously reduced cell number via accumulation in G0-G1 phase of the cell cycle in the absence of notablely increased apoptosis in colon cancer cell lines HT29 and SW620 (S phase arrest in HCT116). Moreover, inhibition of p110β expression increased oxaliplatin-induced cell apoptosis and cell cycle arrest in HT29, HCT116 and SW620 cell lines. In addition, increases of cleaved caspase-3 and cleaved PARP induced by oxaliplatin treatment were determined by immunoblotting in p110β knockdown group compared with normal control group and wildtype group. It is concluded that down-regulated expression of p110β could inhibit colon cancer cells proliferation and result in increased chemosensitivity of colorectal cancer cells to oxaliplatin through augmentation of oxaliplatin-induced cell apoptosis and cell cycle arrest.展开更多
文摘目的:检测大肠癌原发灶与淋巴结(lymph node,LN)转移灶中磷脂酰肌醇3-激酶(phosphatidylinositol 3-kinase,PI3K)p110α、PI3Kp110β与B细胞淋巴瘤基因-2(B c e l l lymphoma 2,Bcl-2)、CyclinD1的表达情况,探讨在大肠癌发生及转移中的相关性及其与临床病理因素及预后的关系.方法:采用免疫组织化学方法在30例正常大肠黏膜组织,52例未发生LN转移的大肠癌组织,50例发生LN转移的大肠癌原发灶及其转移灶中检测PI3Kp110α、PI3Kp110β、Bcl-2和CyclinD1的表达情况及其差异,分析PI3Kp110α、PI3Kp110β与Bcl-2和CyclinD1之间的相关性以及与临床病理因素及其预后的关系.结果:(1)PI3Kp110α与Bcl-2在无LN转移组、有LN转移组的原发灶和其相应LN转移灶中的表达均高于正常肠黏膜组(P<0.05);PI3Kp110β与CyclinD1在无LN转移组、有LN转移组的原发灶和其相应LN转移灶中的表达均高于正常肠黏膜组,且在有LN转移的原发灶中的表达均高于其相应LN转移灶和无LN转移组肠癌中的表达(P<0.05);(2)PI3Kp110α与Bcl-2在4组中均呈正相关(P<0.05);PI3Kp110α与CyclinD1在正常肠黏膜组,无LN转移组和有LN转移组中均呈正相关(P<0.05);P I3Kp110β与Bcl-2和CyclinD1在4组中均呈正相关(P<0.05);(3)PI3Kp110α、PI3Kp110β和CyclinD1蛋白的表达与肿瘤分化程度及LN转移相关,Bcl-2的表达与肿瘤分化程度相关;(4)Kaplan-Meier分析显示:PI3Kp110α、PI3Kp110β、Bcl-2和CyclinD1为影响大肠癌预后的因素之一;Cox比例风险模型分析显示:PI3Kp110α、PI3Kp110β是影响大肠癌患者预后的独立因素.结论:(1)PI3Kp110α、PI3Kp110β与Bcl-2、CyclinD1在大肠癌原发灶及转移灶中的表达均高于正常黏膜,在肿瘤的发生发展中发挥重要作用;(2)大肠癌LN转移灶中,PI3Kp110α与PI3Kp110β分别通过影响Bcl-2及CyclinD1对转移灶中的肿瘤生长起促进作用;(3)PI3Kp110α、PI3Kp110β、Bcl-2和CyclinD1与大肠癌的分化程度有关,且PI3Kp110α、PI3Kp110β和CyclinD1与大肠癌的转移密切相关;(4)PI3Kp110α和PI3Kp110β是影响大肠癌预后的独立危险因素.
基金supported by grants from National Natural Science Foundation of China(No.81072431,30872472,30973496,and30800569)a grant from Innovation Founda-tion of Huazhong University of Science and Technology(No.2010MS027)+1 种基金grants of Foundation of Program973(No.2009CB521802)Special Fund for Central University Basic Scientific Research(No.2011JC062,and2011JC063)
文摘This study examined the synergetic effect of class ⅠA Phosphoinositide 3-kinases catalytic subunit p110β knockdown in conjunction with oxaliplatin treatment on colon cancer cells. Down-regulation of p110β by siRNA interference and oxaliplatin treatment were applied in colon cancer cell lines HT29, SW620 and HCT116. MTT assay was used to measure the inhibitory effect of p110β knockdown on the proliferation of colon cancer cell lines. SubG1 assay and Annexin-Ⅴ FITC/PI double-labeling cytometry were applied to detect cell apoptosis. And cell cycle was evaluated by using PI staining and flow cytometry. The expression of caspase 3, cleaved PARP, p-Akt, T-Akt and p110β was determined by western blotting. The results suggested that down-regulation of p110β expression by siRNA obviously reduced cell number via accumulation in G0-G1 phase of the cell cycle in the absence of notablely increased apoptosis in colon cancer cell lines HT29 and SW620 (S phase arrest in HCT116). Moreover, inhibition of p110β expression increased oxaliplatin-induced cell apoptosis and cell cycle arrest in HT29, HCT116 and SW620 cell lines. In addition, increases of cleaved caspase-3 and cleaved PARP induced by oxaliplatin treatment were determined by immunoblotting in p110β knockdown group compared with normal control group and wildtype group. It is concluded that down-regulated expression of p110β could inhibit colon cancer cells proliferation and result in increased chemosensitivity of colorectal cancer cells to oxaliplatin through augmentation of oxaliplatin-induced cell apoptosis and cell cycle arrest.
