CAJ | 학술논문

目的筛选与斜坡脊索瘤骨侵袭程度相关的蛋白.方法收集起源于蝶岩交界区斜坡脊索瘤患者的肿瘤标本,共35例.12例行同位素标记相对和绝对定量(iTRAQ)蛋白质组学试验(试验组),23例行免疫组化染色验证(验证组).依照骨侵袭程度分型,试验组中膨胀内生型(Ⅰ型)和外生型(Ⅱ型)各6例;验证组Ⅰ和Ⅱ型分别有10例和13例.试验组应用iTRAQ法行差异蛋白检测,应用Panther软件行Gene Ontology功能分析,应用IPA软件行蛋白通路与网络分析.结果共2 251个蛋白获得定量鉴定结果,Ⅰ、Ⅱ型斜坡脊索瘤有250个蛋白存在差异表达,与Ⅱ型相比,Ⅰ型59个蛋白表达上调,191个蛋白下调.Ⅰ和Ⅱ型脊索瘤细胞在哺乳动物雷帕霉素靶蛋白(mTOR)通路表达水平差异有统计学意义(P＜0.01),Ⅰ型的炎性活性相关蛋白表达增强,细胞外基质蛋白和细胞骨架蛋白表达水平低于Ⅱ型.分子网络分析提示,可以调控细胞基质蛋白和骨架蛋白表达的转化生长因子β1(TGFβ1)表达水平降低.免疫组化显示,验证组中TGFβ1、第10号染色体缺失的磷酸酶和张力蛋白同源等位基因(PTEN)的表达阳性率分别为95.6％ (22/23)、69.6％ (16/23),与Ⅰ型比较,Ⅱ型脊索瘤TGFβ1、PTEN的表达强度高(P =0.033,P=0.004);验证组中mTOR的阳性表达率为80.7％ (20/23),Ⅰ和Ⅱ型间mTOR的阳性表达率差异无统计学意义(P=0.092).结论 TGFβ1可能在斜坡脊索瘤的骨侵袭过程中发挥重要作用,机制可能与其介导炎性细胞反应增高及细胞骨架蛋白表达下降有关.PI3K-AKt-mTOR信号通道可能与脊索瘤的骨侵袭发生机制有关,但mTOR蛋白可能与脊索瘤的骨侵袭程度无关.PTEN的表达水平可能与脊索瘤的骨侵袭程度有关.
목적 사선여사파척색류골침습정도상관적단백.방법 수집기원우접암교계구사파척색류환자적종류표본,공35례.12례행동위소표기상대화절대정량(iTRAQ)단백질조학시험(시험조),23례행면역조화염색험증(험증조).의조골침습정도분형,시험조중팽창내생형(Ⅰ형)화외생형(Ⅱ형)각6례;험증조Ⅰ화Ⅱ형분별유10례화13례.시험조응용iTRAQ법행차이단백검측,응용Panther연건행Gene Ontology공능분석,응용IPA연건행단백통로여망락분석.결과 공2 251개단백획득정량감정결과,Ⅰ、Ⅱ형사파척색류유250개단백존재차이표체,여Ⅱ형상비,Ⅰ형59개단백표체상조,191개단백하조.Ⅰ화Ⅱ형척색류세포재포유동물뢰파매소파단백(mTOR)통로표체수평차이유통계학의의(P＜0.01),Ⅰ형적염성활성상관단백표체증강,세포외기질단백화세포골가단백표체수평저우Ⅱ형.분자망락분석제시,가이조공세포기질단백화골가단백표체적전화생장인자β1(TGFβ1)표체수평강저.면역조화현시,험증조중TGFβ1、제10호염색체결실적린산매화장력단백동원등위기인(PTEN)적표체양성솔분별위95.6％ (22/23)、69.6％ (16/23),여Ⅰ형비교,Ⅱ형척색류TGFβ1、PTEN적표체강도고(P =0.033,P=0.004);험증조중mTOR적양성표체솔위80.7％ (20/23),Ⅰ화Ⅱ형간mTOR적양성표체솔차이무통계학의의(P=0.092).결론 TGFβ1가능재사파척색류적골침습과정중발휘중요작용,궤제가능여기개도염성세포반응증고급세포골가단백표체하강유관.PI3K-AKt-mTOR신호통도가능여척색류적골침습발생궤제유관,단mTOR단백가능여척색류적골침습정도무관.PTEN적표체수평가능여척색류적골침습정도유관.
Objective To screen the proteins associated with the degree of bone invasion in clivus chordomas.Methods A total of 35 tumor specimens of patients with clivus chordoma originated from the spheno petrosal junction were collected.Twelve specimens of isobaric tags for relative and absolute quantitation (iTRAQ) proteomics experiment (experimental group) were performed,23 were validated by immunohistochemical staining (validated group).According to the classification criteria of bone invasive degree,they were classified into endophytic type (Type Ⅰ) or exophytic type (Type Ⅱ) (n =6 in each group) in the experimental group.Type Ⅰ and type Ⅱ in the validated group were 10 and 13 specimens respectively.The iTRAQ technique was used for the detection of the difference proteins.Panther software was used for functional analysis of gene ontology (GO).IPA software was used for protein pathway and network analysis.Results A total of 2251 proteins obtained quantitative identification results,250 proteins had difference expression in type Ⅰ and type Ⅱ clivus chordomas.Compared with type Ⅱ,59 proteins were up-regulated and 191 were down-regulated in type Ⅰ.There was significant difference in the expression levels of chordomas cells of the mammalian target of rapamycin (mTOR) pathway between the type Ⅰ and type Ⅱ chordoma cells (P ＜0.01).The inflammatory activity-associated protein expression of type Ⅰ enhanced.The expression levels of extracellular matrix proteins and cytoskeletal proteins were lower than those of type Ⅱ.Molecular network analysis suggested that the expression levels of transforming growth factor beta 1 (TGFβ31) of regulating the cell matrix protein and cytoskeleton protein expression were low.Immunohistochemistry showed that the expression positive rates of TGFβ1,phosphatase and tensin homolog deleted on chromosome ten (PTEN) in the validated group were 95.6％ (22/23) and 69.6％ (16/23).Compare with type Ⅰ,the expression intensity of type Ⅱ chordoma TGFβ1 and PTEN was high (P =0.033,P =0.004);the positive expression rate of mTOR in the validated group was 80.7％ (20/23).There was no significant difference in the positive expression rate between the type Ⅰ and the type Ⅱ (P =0.092).Conclusions TGFβ1 may play an important role in bone invasion of upper-middle clivus chordoma.The mechanisms may be associated with its mediating increased inflammatory cell response and the decreased expression of cytoskeletal proteins.The PI3K-AKt-mTOR signal pathway may be associated with the pathogenesis of bone invasion of chordoma,however,mTOR protein itself may not be associated with the bone invasion degree of chordoma.The expression level of PTEN may be associated with the bone invasion degree of chordoma.