CAJ | 학술논문

背景与目的:肺癌易转移至骨并导致溶骨性破坏的具体机制目前还不清楚,本研究观察CCN3在肺癌骨转移灶中的表达水平以及在前体成骨细胞(MC3T3-E1)与前体破骨细胞(RAW264.7)共培养体系研究CCN3重组蛋白对前体破骨细胞分化的影响.方法:采用免疫组化检测9例肺癌骨转移转灶组织及相应癌旁组织中CCN3的表达;构建RAW264.7细胞与MC3T3-E1细胞共培养模型,向共培养体系中加或不加800?ng/mL的CCN3重组蛋白,采用Real-time? PCR检测RAW264.7细胞的破骨细胞标志基因抗酒石酸酸性磷酸酶(tartrate-resistant?acid? phosphatase,TRAP)、组织蛋白酶K(cathepsin? K,CK)、降钙素受体(calcitonin? receptor,CTR) mRNA的表达,并通过TRAP染色法对破骨细胞进行鉴定.结果:肺癌骨转移灶组织中的CCN3表达量明显高于癌旁组织;CCN3重组蛋白可显著促进共培养体系中的RAW264.7细胞的破骨细胞标志基因TRAP、CK、CTR的mRNA的表达量;TRAP染色可见对照组TRAP阳性细胞数[(6.2±1.48)个]明显低于加了CCN3重组蛋白的试验组[(21.4±3.04)个],差异具有统计学意义(P<0.05).结论:CCN3在肺癌骨转移灶中高表达及其可诱导破骨细胞分化,CCN3可能在肺癌骨转移和溶骨性破坏中具有重要作用.
배경여목적:폐암역전이지골병도치용골성파배적구체궤제목전환불청초,본연구관찰CCN3재폐암골전이조중적표체수평이급재전체성골세포(MC3T3-E1)여전체파골세포(RAW264.7)공배양체계연구CCN3중조단백대전체파골세포분화적영향.방법:채용면역조화검측9례폐암골전이전조조직급상응암방조직중CCN3적표체;구건RAW264.7세포여MC3T3-E1세포공배양모형,향공배양체계중가혹불가800?ng/mL적CCN3중조단백,채용Real-time? PCR검측RAW264.7세포적파골세포표지기인항주석산산성린산매(tartrate-resistant?acid? phosphatase,TRAP)、조직단백매K(cathepsin? K,CK)、강개소수체(calcitonin? receptor,CTR) mRNA적표체,병통과TRAP염색법대파골세포진행감정.결과:폐암골전이조조직중적CCN3표체량명현고우암방조직;CCN3중조단백가현저촉진공배양체계중적RAW264.7세포적파골세포표지기인TRAP、CK、CTR적mRNA적표체량;TRAP염색가견대조조TRAP양성세포수[(6.2±1.48)개]명현저우가료CCN3중조단백적시험조[(21.4±3.04)개],차이구유통계학의의(P<0.05).결론:CCN3재폐암골전이조중고표체급기가유도파골세포분화,CCN3가능재폐암골전이화용골성파배중구유중요작용.
? Background and purpose:Lung cancer frequently metastasizes to bone resulting in osteolytic lesions with unknown mechanisms. This study aimed to investigate CCN3 expressed in bone metastatic lesions from lung cancer, and determine the effects of CCN3 on the differentiation of RAW264.7 cells which co-culture with MC3T3-E1 cells.? Methods:Immunohistochemical staining was used to detect the expression of CCN3 in bone metastatic lesions and it’s adjacent tissues; Using a RAW264.7 and MC3T3-E1 cell co-culture system, MC3T3-E1 cells and RAW264.7 cells were exposed to recombination CCN3 (800 ng/mL) in a co-culture system. To examine osteoclast differentiation, the expression of osteoclastic markers tartrate-resistant acid phosphatase (TRAP), cathepsin K (CK) and calcitonin receptor (CTR) mRNA analyzed by Real time-PCR, and osteoclast formation were determined by tartrate-resistant acid phosphatase staining. Results:The expression of CCN3 in bone metastatic lesions were significantly higher than that in the adjacent tissues; CCN3 increased the expression of osteoclast marker genes mRNA in RAW264.7 cocultured with MC3T3-E1; Recombination CCN3 significantly stimulated the formation of osteoclast (21.4±3.04) in co-culture system compared with the control (6.2±1.48), a statistically significant difference (P<0.05). Conclusion:Our data demonstrate CCN3 might play an important role in lung cancer metastasizes to bone and to promote osteolytic lesions.