CAJ | 학술논문

背景与目的：胃癌是我国常见的消化道恶性肿瘤，术后易复发和转移。前期研究发现，蛋白酪氨酸磷酸酶1B(protein tyrosine phosphatase 1B，PTP1B)基因与胃癌肿瘤大小、淋巴结转移及TNM分期有关。本研究通过RNA干扰技术和基因克隆技术分别沉默和上调胃癌细胞中PTP1B基因的表达，观察PTP1B基因对胃癌细胞增殖和迁移能力的影响。方法：将靶向沉默PTP1B基因的短发夹RNA(short hairpin RNA，shRNA)序列和克隆人的PTP1B cDNA基因序列分别转染MKN28和MKN45细胞，采用实时定量PCR(quantitative real-time PCR，qRT-PCR)、蛋白[质]印迹法(Western blot)分别检测转染后细胞中PTP1B基因和蛋白的表达水平，使用细胞计数试剂盒(cell counting kit-8，CCK-8)、Transwell迁移试验和划痕试验分别观察PTP1B基因对细胞增殖和迁移能力的影响。结果：转染shRNA后，MKN28细胞中PTP1B mRNA和蛋白表达量与空白对照组、阴性对照组相比抑制显著(P＜0.05)。CCK-8增殖活性实验显示，shRNA沉默PTP1B基因表达后能显著抑制胃癌MKN28细胞在48、72和96 h的增殖活性(P＜0.05)。Transwell迁移试验和划痕实验显示，PTP1B表达下调后胃癌MKN28细胞的迁移能力受到显著抑制(P＜0.05)。而提高PTP1B在MKN45细胞中表达后，细胞的增殖和迁移能力则显著提高(P＜0.05)。结论：PTP1B基因是胃癌细胞增殖和迁移的重要调控因子。
배경여목적：위암시아국상견적소화도악성종류，술후역복발화전이。전기연구발현，단백락안산린산매1B(protein tyrosine phosphatase 1B，PTP1B)기인여위암종류대소、림파결전이급TNM분기유관。본연구통과RNA간우기술화기인극륭기술분별침묵화상조위암세포중PTP1B기인적표체，관찰PTP1B기인대위암세포증식화천이능력적영향。방법：장파향침묵PTP1B기인적단발협RNA(short hairpin RNA，shRNA)서렬화극륭인적PTP1B cDNA기인서렬분별전염MKN28화MKN45세포，채용실시정량PCR(quantitative real-time PCR，qRT-PCR)、단백[질]인적법(Western blot)분별검측전염후세포중PTP1B기인화단백적표체수평，사용세포계수시제합(cell counting kit-8，CCK-8)、Transwell천이시험화화흔시험분별관찰PTP1B기인대세포증식화천이능력적영향。결과：전염shRNA후，MKN28세포중PTP1B mRNA화단백표체량여공백대조조、음성대조조상비억제현저(P＜0.05)。CCK-8증식활성실험현시，shRNA침묵PTP1B기인표체후능현저억제위암MKN28세포재48、72화96 h적증식활성(P＜0.05)。Transwell천이시험화화흔실험현시，PTP1B표체하조후위암MKN28세포적천이능력수도현저억제(P＜0.05)。이제고PTP1B재MKN45세포중표체후，세포적증식화천이능력칙현저제고(P＜0.05)。결론：PTP1B기인시위암세포증식화천이적중요조공인자。
Background and purpose:Gastric cancer is the most common malignant tumor of digestive tract, and the possibility of postoperative recurrence and metastasis is higher. Our previous studies showed that protein tyrosine phosphatase1B (PTP1B) gene is closely correlated with tumor size, lymph node metastasis and TNM stage of gastric cancer. The purpose of the present study was to explore the effect ofPTP1B gene on cell proliferation and migration of gastric cancer cell lines.Methods:Short hairpin RNA (shRNA) sequence targetingPTP1B gene and PTP1B cDNA were transfected into MKN28 and MKN45 cells, respectively. The expression of PTP1B mRNA and its protein in MKN28 and MKN45 cells were detected by quantitative real-time PCR (qRT-PCR) and Western blot, respectively. The effect of PTP1B on cell proliferation and migration was respectively assessed by cell counting kit-8 (CCK-8) assay, Transwell migration assay and wound healing assay.Results:Compared with blank and negative controls, the expressions of PTP1B mRNA and protein in MKN28 cells were successfully suppressed after the cells were transfected with shRNA (P<0.05). As CCK-8 test showed, the proliferation of MKN28 cells was successfully restrained at 48, 72 and 96 h after transfected with shRNA compared with blank control and negative control (P<0.05). Transwell and wound healing test were performed after PTP1B expression was interfered by shRNA. The result demonstrated that migration of MKN28 cells was signiifcantly inhibited (P<0.05). On the contrary, the expressions of PTP1B mRNA and protein in MKN45 cells were obviously enhanced after the cells were transfected with PTP1B cDNA. And the proliferation and migration of cells were significantly increased.Conclusion:PTP1B gene is an important enchancer for the proliferation and migration of gastric cancer.