胃癌组织中LIN28B和c-myc的表达及其临床意义
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摘要
目的检测胃癌组织中LIN28B和c-myc的表达,分析其临床意义,并初步探讨其可能的作用机制。方法应用免疫组化SP法检测90例胃癌和30例癌旁正常胃黏膜石蜡组织中LIN28B和c-myc蛋白的表达水平;应用实时荧光定量PCR方法和Western blotting法检测23例新鲜胃癌组织及相应癌旁正常组织中LIN28B和c-myc mRNA及蛋白的表达量。结果胃癌组织中LIN28B mRNA(△Ct:6.33±2.54)及蛋白(0.601±0.22)的表达水平显著高于癌旁正常组织(△Ct:1.05±0.64,0.124±0.088,P<0.05);LIN28B在胃癌组织中表达阳性率(76.7%,69/90)较癌旁正常胃黏膜石蜡组织中阳性率(66.7%,20/30)高,差异有统计学意义(H=4.195,P<0.05);c-myc在胃癌组织中表达阳性率(81.1%,73/90)较正常胃黏膜石蜡组织中阳性率(60.0%,18/30)高,差异有统计学意义(H=3.887,P<0.05);LIN28B蛋白表达与胃癌分化程度、淋巴结转移、浸润深度、临床分期有相关性(P<0.05),与年龄、性别、肿瘤大小、部位无相关性(P>0.05);LIN28B与c-myc在胃癌组织中的表达有显著相关性(r=0.252,P<0.05)。结论LIN28B和c-myc基因的表达及相互作用与胃癌的发生、发展及生物学行为密切相关,对判断预后有参考作用,其机制可能与胃癌干细胞及LIN28B/let-7/c-myc信号环路有关。
Objective To measure the expression of LIN28 B and c-myc in gastric cancer and to determine their association with the clinicopathological characteristics and prognosis of patients with gastric cancer. Methods The expression of LIN28 B and c-myc mRNA and protein in 23 fresh gastric cancer tissues and adjacent normal tissues were measured by real-time PCR and Western blotting analysis. The expression levels of LIN28 B and c-myc protein in 90 paraffin blocks of gastric cancer and 30 adjacent normal gastric tissues were determined by substance P immunohistochemical technique. Results The LIN28 B mRNA( △Ct: 6. 33 ± 2. 54) and protein(0. 601 ± 0. 22) in fresh gastric cancer tissues were significantly higher than those in adjacent normal tissues( △Ct: 1. 05 ± 0. 64,0. 124 ± 0. 088,P < 0. 05).LIN28 B protein was positive in 76. 7% of the paraffin blocks of gastric cancer and in 66. 7% of normal gastric tissues,the difference was statistically significant( H = 4. 195,P < 0. 05). c-myc protein was positive in 81. 1% of the paraffin blocks of gastric cancer and in 60% of normal gastric tissues,the difference was statistically significant( H = 3. 887,P< 0. 05). LIN28 B protein was significantly correlated with tumor differentiation,lymphatic metastasis,invasive depth,and clinical stage( P < 0. 05),but not correlated with age,gender,tumor size,and tumor areas( P > 0. 05). The fiveyear survival rate of LIN28B-positive patients was significantly lower than that of LIN28B-negative patients( P < 0. 05).The five-year survival rates of c-myc-positive patients were significantly lower than those of c-myc-negative patients( P <0. 05). The expression of LIN28 B and c-myc in gastric cancer tissue had significant correlation( r = 0. 252,P < 0. 05).Conclusion LIN28 B and c-myc are highly correlated with the biological behavior of gastric carcinoma and thus facilitate the evaluation of gastric carcinoma prognosis. The mechanism may be related to gastric cancer stem cells and the circuits conduction path of LIN28 B / let-7 / c-myc.
Objective To measure the expression of LIN28 B and c-myc in gastric cancer and to determine their association with the clinicopathological characteristics and prognosis of patients with gastric cancer. Methods The expression of LIN28 B and c-myc mRNA and protein in 23 fresh gastric cancer tissues and adjacent normal tissues were measured by real-time PCR and Western blotting analysis. The expression levels of LIN28 B and c-myc protein in 90 paraffin blocks of gastric cancer and 30 adjacent normal gastric tissues were determined by substance P immunohistochemical technique. Results The LIN28 B mRNA( △Ct: 6. 33 ± 2. 54) and protein(0. 601 ± 0. 22) in fresh gastric cancer tissues were significantly higher than those in adjacent normal tissues( △Ct: 1. 05 ± 0. 64,0. 124 ± 0. 088,P < 0. 05).LIN28 B protein was positive in 76. 7% of the paraffin blocks of gastric cancer and in 66. 7% of normal gastric tissues,the difference was statistically significant( H = 4. 195,P < 0. 05). c-myc protein was positive in 81. 1% of the paraffin blocks of gastric cancer and in 60% of normal gastric tissues,the difference was statistically significant( H = 3. 887,P< 0. 05). LIN28 B protein was significantly correlated with tumor differentiation,lymphatic metastasis,invasive depth,and clinical stage( P < 0. 05),but not correlated with age,gender,tumor size,and tumor areas( P > 0. 05). The fiveyear survival rate of LIN28B-positive patients was significantly lower than that of LIN28B-negative patients( P < 0. 05).The five-year survival rates of c-myc-positive patients were significantly lower than those of c-myc-negative patients( P <0. 05). The expression of LIN28 B and c-myc in gastric cancer tissue had significant correlation( r = 0. 252,P < 0. 05).Conclusion LIN28 B and c-myc are highly correlated with the biological behavior of gastric carcinoma and thus facilitate the evaluation of gastric carcinoma prognosis. The mechanism may be related to gastric cancer stem cells and the circuits conduction path of LIN28 B / let-7 / c-myc.
引文
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[11]Shimizu S,Takehara T,Hikita H,et al.The let-7 family of microRNAs inhibits Bcl-x L expression and potentiates sorafenib-induced apoptosis in human hepatocellular carcinoma[J].J Hepatol,2010,52(5):698-704.
[12]Dong Q,Meng P,Wang T,et al.MicroRNA let-7a inhibits proliferation of human prostate cancer cells in vitro and in vivo by targeting E2F2 and CCND2[J].PLo S One,2010,5(4):e10147.
[13]Leppert U,Henke W,Huang X,et al.Post-transcriptional fine-tuning of COP9 signalosome subunit biosynthesis is regulated by the cMyc/Lin28B/let-7 pathway[J].J Mol Biol,2011,409(5):710-721.
[14]Mc Carty MF.Metformin may antagonize Lin28 and/or Lin28B activity,thereby boosting let-7 levels and antagonizing cancer progression[J].Med Hypotheses,2012,78(2):262-269.
[15]Thornton JE,Gregory RI.How does Lin28 let-7 control development and disease?[J].Trends Cell Biol,2012,22(9):474-482.
[2]Fu X,Meng Z,Liang W,et al.miR-26a enhances miRNA biogenesis by targeting Lin28B and Zcchc11 to suppress tumor growth and metastasis[J].Oncogene,2014,33(34):4296-4306.
[3]Chang TC,Zeitels LR,Hwang HW,et al.Lin-28B transactivation is necessary for Myc-mediated let-7 repression and proliferation[J].Proc Natl Acad Sci U S A,2009,106(9):3384-3389.
[4]Wang JM,Lin SR.Progress in the clinical research of gastric cancer[J].Chin J Gastroenterol Hepatol,2012,21(1):3-5.王婕敏,林三仁.胃癌研究及诊治新进展[J].胃肠病学和肝病学杂志,2012,21(1):3-5.
[5]Zhou J,Ng SB,Chng WJ.LIN28/LIN28B:an emerging oncogenic driver in cancer stem cells[J].Int J Biochem Cell Biol,2013,45(5):973-978.
[6]King CE,Cuatrecasas M,Castells A,et al.LIN28B promotes colon cancer progression and metastasis[J].Cancer Res,2011,71(12):4260-4268.
[7]Permuth-Wey J,Kim D,Tsai YY,et al.LIN28B polymorphisms influence susceptibility to epithelial ovarian cancer[J].Cancer Res,2011,71(11):3896-3903.
[8]Hamano R,Miyata H,Yamasaki M,et al.High expression of Lin28 is associated with tumour aggressiveness and poor prognosis of patients in oesophagus cancer[J].Br J Cancer,2012,106(8):1415-1423.
[9]Cai WY,Wei TZ,Luo QC,et al.The Wnt-β-catenin pathway represses let-7 microRNA expression through transactivation of Lin28 to augment breast cancer stem cell expansion[J].J Cell Sci,2013,126(Pt13):2877-2889.
[10]Thornton JE,Gregory RI.How does Lin28 let-7 control development and disease?[J].Trends Cell Biol,2012,22(9):474-482.
[11]Shimizu S,Takehara T,Hikita H,et al.The let-7 family of microRNAs inhibits Bcl-x L expression and potentiates sorafenib-induced apoptosis in human hepatocellular carcinoma[J].J Hepatol,2010,52(5):698-704.
[12]Dong Q,Meng P,Wang T,et al.MicroRNA let-7a inhibits proliferation of human prostate cancer cells in vitro and in vivo by targeting E2F2 and CCND2[J].PLo S One,2010,5(4):e10147.
[13]Leppert U,Henke W,Huang X,et al.Post-transcriptional fine-tuning of COP9 signalosome subunit biosynthesis is regulated by the cMyc/Lin28B/let-7 pathway[J].J Mol Biol,2011,409(5):710-721.
[14]Mc Carty MF.Metformin may antagonize Lin28 and/or Lin28B activity,thereby boosting let-7 levels and antagonizing cancer progression[J].Med Hypotheses,2012,78(2):262-269.
[15]Thornton JE,Gregory RI.How does Lin28 let-7 control development and disease?[J].Trends Cell Biol,2012,22(9):474-482.