中华检验医学杂志
中華檢驗醫學雜誌
중화검험의학잡지
CHINESE JOURNAL OF LABORATORY MEDICINE
2011年
9期
830-835
,共6页
郭飞%陈玉华%刘永霞%朱小丽%陈伟%胡王强%陈占国%沈默%陶志华
郭飛%陳玉華%劉永霞%硃小麗%陳偉%鬍王彊%陳佔國%瀋默%陶誌華
곽비%진옥화%류영하%주소려%진위%호왕강%진점국%침묵%도지화
膀胱肿瘤%受体蛋白质酪氨酸激酶类%选择性剪接
膀胱腫瘤%受體蛋白質酪氨痠激酶類%選擇性剪接
방광종류%수체단백질락안산격매류%선택성전접
Urinary bladder neoplasms%Receptor protein-tyrosine kinases%Alternative splicing
目的 探讨RON mRNA及其变异体在膀胱肿瘤组织中的表达与临床意义。方法选择63例膀胱移行上皮癌(TCCB)、7例膀胱内翻性乳头状瘤(IPB)、9例膀胱低度恶性潜能尿路上皮瘤(PUNLMP)患者和12例外伤性膀胱破裂且经病理证实无肿瘤细胞的正常膀胱黏膜组织患者。其中,病理Ⅰ、Ⅱ和Ⅲ级患者分别为30、15和18例,临床Tis+ T1和T2 +T3 +T4期患者分别为44例和15例;用实时荧光定量RT-PCR检测其RON mRNA相对表达量,以GAPDH mRNA为内标物,用RONmRNA/GAPDH mRNA比值表示RON mRNA相对表达量;用RT-PCR检测RON mRNA选择性剪切形成的变异体;用测序检测PCR产物中可能存在的变异体,并分析变异体在不同组织间及TCCB中不同病理分级及临床分期间阳性表达率的差异。结果RON mRNA在TCCB、IPB、PUNLMP及正常膀胱黏膜组织中均见阳性表达,其RON mRNA/GAPDH mRNA比值分别为4.9×10-3(1.8×10-3 ~1.0× 10-2)、3.8×10-3(2.4×10-3~1.7×10-2)、4.9×10-3(1.7×10-3~1.1 ×10-2)、1.0×10-3(4.5×10-4 ~2.8×10-3),且不同组织间的表达差异均有统计学意义(x2K-W= 17.278,P<0.05);RON mRNA/GAPDH mRNA比值在TCCB病理Ⅰ、Ⅱ、Ⅲ级分别为3.7×10-3(1.3×10-3 ~7.5×10-3)、4.9×10-3(1.9X10-3~1.1 ×10-2)、8.9×10-3(2.7×10-3~8.0×10-2),且差异有统计学意义(x2K-W=7.341,P<0.05);RON mRNA/GAPDH mRNA比值在TCCB临床Tis+ T1、T2 +T3 +T4期分别为3.5×10-3(1.2×10-3 ~7.7×10-3)、9.7× 10-3(2.9×10-3~5.3 ×10-2),差异也有统计学意义(Z= -2.306,P<0.05)。但正常膀胱黏膜组未发现RON mRNA变异体,而在膀胱肿瘤组织中外显子11剪切缺失(E11△)的阳性表达率为70%( 55/79)。在TCCB、IPB、PUNLMP中E11△阳性表达率分别为71% (45/63)、57% (4/7)、67% (6/9),而不同病理组织中的E11△阳性表达率差异无统计学意义(x2=0.620,P>0.05);在不同TCCB病理分级和临床分期中其阳性率差异亦无统计学意义(Z值分别为0.221、0.538,P均>0.05)。发现1种正常黏膜组织中没有的新变异体,即RON基因外显子的3 476~3 539 bp剪切缺失形成的变异体(E3 476 ~3 539△)。膀胱肿瘤组织中E3 476~3 539△的阳性表达率为56% (44/79),在TCCB、IPB、PUNLMP中E 3 476 ~3 539△的阳性表达率分别为57%( 36/63)、43% (3/7)、56% (5/9),但各病理组织间的阳性率差异无统计学意义(x2=0.517,P>0.05);在TCCB不同病理分级和临床分期中其阳性率分别为40% (12/30)、67% (10/15)、78% (14/18)、48%(21/44)、80% (12/15),差异有统计学意义(Z值分别为7.285、5.041,P均<0.05)。结论 RONmRNA的表达与TCCB病理分级及临床分期相关,RON可能在TCCB发生发展的过程中发挥重要作用;在正常膀胱黏膜中未见RON mRNA剪切形成的变异体,而在膀胱肿瘤组织中都有不同程度的表达;E 3 476~3 539△的表达与TCCB病理分级及临床分期相关,RON mRNA变异体可能参与了膀胱肿瘤的发生。
目的 探討RON mRNA及其變異體在膀胱腫瘤組織中的錶達與臨床意義。方法選擇63例膀胱移行上皮癌(TCCB)、7例膀胱內翻性乳頭狀瘤(IPB)、9例膀胱低度噁性潛能尿路上皮瘤(PUNLMP)患者和12例外傷性膀胱破裂且經病理證實無腫瘤細胞的正常膀胱黏膜組織患者。其中,病理Ⅰ、Ⅱ和Ⅲ級患者分彆為30、15和18例,臨床Tis+ T1和T2 +T3 +T4期患者分彆為44例和15例;用實時熒光定量RT-PCR檢測其RON mRNA相對錶達量,以GAPDH mRNA為內標物,用RONmRNA/GAPDH mRNA比值錶示RON mRNA相對錶達量;用RT-PCR檢測RON mRNA選擇性剪切形成的變異體;用測序檢測PCR產物中可能存在的變異體,併分析變異體在不同組織間及TCCB中不同病理分級及臨床分期間暘性錶達率的差異。結果RON mRNA在TCCB、IPB、PUNLMP及正常膀胱黏膜組織中均見暘性錶達,其RON mRNA/GAPDH mRNA比值分彆為4.9×10-3(1.8×10-3 ~1.0× 10-2)、3.8×10-3(2.4×10-3~1.7×10-2)、4.9×10-3(1.7×10-3~1.1 ×10-2)、1.0×10-3(4.5×10-4 ~2.8×10-3),且不同組織間的錶達差異均有統計學意義(x2K-W= 17.278,P<0.05);RON mRNA/GAPDH mRNA比值在TCCB病理Ⅰ、Ⅱ、Ⅲ級分彆為3.7×10-3(1.3×10-3 ~7.5×10-3)、4.9×10-3(1.9X10-3~1.1 ×10-2)、8.9×10-3(2.7×10-3~8.0×10-2),且差異有統計學意義(x2K-W=7.341,P<0.05);RON mRNA/GAPDH mRNA比值在TCCB臨床Tis+ T1、T2 +T3 +T4期分彆為3.5×10-3(1.2×10-3 ~7.7×10-3)、9.7× 10-3(2.9×10-3~5.3 ×10-2),差異也有統計學意義(Z= -2.306,P<0.05)。但正常膀胱黏膜組未髮現RON mRNA變異體,而在膀胱腫瘤組織中外顯子11剪切缺失(E11△)的暘性錶達率為70%( 55/79)。在TCCB、IPB、PUNLMP中E11△暘性錶達率分彆為71% (45/63)、57% (4/7)、67% (6/9),而不同病理組織中的E11△暘性錶達率差異無統計學意義(x2=0.620,P>0.05);在不同TCCB病理分級和臨床分期中其暘性率差異亦無統計學意義(Z值分彆為0.221、0.538,P均>0.05)。髮現1種正常黏膜組織中沒有的新變異體,即RON基因外顯子的3 476~3 539 bp剪切缺失形成的變異體(E3 476 ~3 539△)。膀胱腫瘤組織中E3 476~3 539△的暘性錶達率為56% (44/79),在TCCB、IPB、PUNLMP中E 3 476 ~3 539△的暘性錶達率分彆為57%( 36/63)、43% (3/7)、56% (5/9),但各病理組織間的暘性率差異無統計學意義(x2=0.517,P>0.05);在TCCB不同病理分級和臨床分期中其暘性率分彆為40% (12/30)、67% (10/15)、78% (14/18)、48%(21/44)、80% (12/15),差異有統計學意義(Z值分彆為7.285、5.041,P均<0.05)。結論 RONmRNA的錶達與TCCB病理分級及臨床分期相關,RON可能在TCCB髮生髮展的過程中髮揮重要作用;在正常膀胱黏膜中未見RON mRNA剪切形成的變異體,而在膀胱腫瘤組織中都有不同程度的錶達;E 3 476~3 539△的錶達與TCCB病理分級及臨床分期相關,RON mRNA變異體可能參與瞭膀胱腫瘤的髮生。
목적 탐토RON mRNA급기변이체재방광종류조직중적표체여림상의의。방법선택63례방광이행상피암(TCCB)、7례방광내번성유두상류(IPB)、9례방광저도악성잠능뇨로상피류(PUNLMP)환자화12예외상성방광파렬차경병리증실무종류세포적정상방광점막조직환자。기중,병리Ⅰ、Ⅱ화Ⅲ급환자분별위30、15화18례,림상Tis+ T1화T2 +T3 +T4기환자분별위44례화15례;용실시형광정량RT-PCR검측기RON mRNA상대표체량,이GAPDH mRNA위내표물,용RONmRNA/GAPDH mRNA비치표시RON mRNA상대표체량;용RT-PCR검측RON mRNA선택성전절형성적변이체;용측서검측PCR산물중가능존재적변이체,병분석변이체재불동조직간급TCCB중불동병리분급급림상분기간양성표체솔적차이。결과RON mRNA재TCCB、IPB、PUNLMP급정상방광점막조직중균견양성표체,기RON mRNA/GAPDH mRNA비치분별위4.9×10-3(1.8×10-3 ~1.0× 10-2)、3.8×10-3(2.4×10-3~1.7×10-2)、4.9×10-3(1.7×10-3~1.1 ×10-2)、1.0×10-3(4.5×10-4 ~2.8×10-3),차불동조직간적표체차이균유통계학의의(x2K-W= 17.278,P<0.05);RON mRNA/GAPDH mRNA비치재TCCB병리Ⅰ、Ⅱ、Ⅲ급분별위3.7×10-3(1.3×10-3 ~7.5×10-3)、4.9×10-3(1.9X10-3~1.1 ×10-2)、8.9×10-3(2.7×10-3~8.0×10-2),차차이유통계학의의(x2K-W=7.341,P<0.05);RON mRNA/GAPDH mRNA비치재TCCB림상Tis+ T1、T2 +T3 +T4기분별위3.5×10-3(1.2×10-3 ~7.7×10-3)、9.7× 10-3(2.9×10-3~5.3 ×10-2),차이야유통계학의의(Z= -2.306,P<0.05)。단정상방광점막조미발현RON mRNA변이체,이재방광종류조직중외현자11전절결실(E11△)적양성표체솔위70%( 55/79)。재TCCB、IPB、PUNLMP중E11△양성표체솔분별위71% (45/63)、57% (4/7)、67% (6/9),이불동병리조직중적E11△양성표체솔차이무통계학의의(x2=0.620,P>0.05);재불동TCCB병리분급화림상분기중기양성솔차이역무통계학의의(Z치분별위0.221、0.538,P균>0.05)。발현1충정상점막조직중몰유적신변이체,즉RON기인외현자적3 476~3 539 bp전절결실형성적변이체(E3 476 ~3 539△)。방광종류조직중E3 476~3 539△적양성표체솔위56% (44/79),재TCCB、IPB、PUNLMP중E 3 476 ~3 539△적양성표체솔분별위57%( 36/63)、43% (3/7)、56% (5/9),단각병리조직간적양성솔차이무통계학의의(x2=0.517,P>0.05);재TCCB불동병리분급화림상분기중기양성솔분별위40% (12/30)、67% (10/15)、78% (14/18)、48%(21/44)、80% (12/15),차이유통계학의의(Z치분별위7.285、5.041,P균<0.05)。결론 RONmRNA적표체여TCCB병리분급급림상분기상관,RON가능재TCCB발생발전적과정중발휘중요작용;재정상방광점막중미견RON mRNA전절형성적변이체,이재방광종류조직중도유불동정도적표체;E 3 476~3 539△적표체여TCCB병리분급급림상분기상관,RON mRNA변이체가능삼여료방광종류적발생。
Objectives To explore the clinical significance of tyrosine kinase receptor RON mRNA expression and it's splicing variant in bladder tumors. Methods Sixty-three cases of transitional cell carcinoma of the bladder (TCCB), including 30 cases of pathological grade I , 15 cases of pathological grade Ⅱ and 18 cases of pathological grade Ⅲ (44 cases of clinical stage Tis + T1, 15 cases of T2 + T3 +T4), 7 inverted papilloma of the bladder ( IPB), 9 cases of papillary urothelial neoplasm of low malignant potential (PUNLMP) and 12 cases of normalbladder mucosa RT-PCR was employed with the internal standard (GAPDHmRNA) to detect the expression of RON mRNA. PCR and direct sequencing was then utilized to identify the potential RON mRNA splicing variants. Finally, the variants' positive rates of expression were analyzed among the different tissues, diverse TCCB pathological grades and clinical stages. Results The expression levels of RON mRNA/GAPDH mRNA among TCCB, IPB, PUNLMP and normal control were 4. 9 × 10-3 ( 1. 8 × 10-3-1.0 × 10-2 ), 3. 8 × 10-3 (2. 4 × 10-3-1.7 × 10-2 ), 4. 9 ×10 -3 ( 1.7 × 10 -3-1.1 × 10 -2 ) and 1.0 × 10-3 (4. 5 × 10-4-2. 8 × 10-3 ) respectively. The difference had a statistical significance (x2K-W = 17. 278 ,P <0. 05 ). The expression levels among pathological grade I, Ⅱ and Ⅲ were 3.7 × 10-3( 1.3 × 10-3-7.5 × 10-3) , 4. 9 × 10-3(1.9 × 10-3-1.1 × 10-2) and 8.9 × 10-3(2. 7 ×10 -3-8.0 × 10 -2 ) respectively. The erpression levels among the clinical stage Tis + T1 and T2 + T3 + T4were 3.5 × 10-3 ( 1.2 × 10 -3-7. 7 × 10-3 ) and 9. 7 × 10 -3 ( 2. 9 × 10-3-5. 3 × 10-2 ). The differences between expression levels were of statistical significance among the different pathological grades ( x2k-W =7. 341, P <0. 05 ) and clinical stages ( Z = - 2. 306, P < 0. 05 ). The positive rates of exon 11deletion(E11△) in TCCB, IPB and PUNLMP were 71% (45/63), 57% (4/7) and 67% (6/9) respectively, andthe total positive rate in bladder tumor tissues was 70%. Meanwhile, expression of the novel RON variant wesnot detected in the normalbladder mucosa. The positive expression rate of E1 1△ has no significant correlationamong the different clinical pathological tissues (x2 = 0. 620, P > 0. 05 ). There was no statistical significancein expression positive rate between different pathological grades ( Z =0. 221, P >0. 05 ) and clinical stages( Z = 0. 538, P > 0. 05) as well. A novel RON splice variant, deletion of RON exon 11 3 476 - 3 539 ( E3476 -3539△) was fond in the pathological tissue. The positive expression rates of E3 476 -3 539 in TCCB,IPB and PUNLMP were 57% (36/63), 43% (3/7) and 56% (5/9) respectively, and the total positive expression rate was 56% (44/79). The positive rates of E3 476 -3 539△ in pathological grade I , Ⅱ and Ⅲ were 40% ( 12/30), 67% (10/15) and 78% (14/18), and it's positive rates in clinical stage Tis +T1and T2 +T3 + T4 were 48% (21/44) and 80% (12/15). The differences in each group had significantly statistical significance ( Z = 7. 285, 5. 041, P < 0. 05 ) . However, the positive rates amongdifferent pathological tissues had no significance (x2 = 0. 517, P > 0. 05 ). Conclusions The expression level of RON mRNA is significantly associated with histological grading and clinical stage. RON may play an important role in the progression ofTCCB. Compared with the normal control, the increased RON variant expression may contribute to the carcinogenesis of the bladder tumor.