中华实验外科杂志
中華實驗外科雜誌
중화실험외과잡지
CHINESE JOURNAL OF EXPERIMENTAL SURGERY
2014年
12期
2729-2731
,共3页
王顺涛%能一全%王君涛%黄韬
王順濤%能一全%王君濤%黃韜
왕순도%능일전%왕군도%황도
WNT10B%癌相关成纤维细胞%乳腺癌
WNT10B%癌相關成纖維細胞%乳腺癌
WNT10B%암상관성섬유세포%유선암
WNT10B%Carcinoma associated fibroblasts%Breast cancer
目的 观察成纤维细胞对乳腺癌细胞MCF-7表达WNT10B和增殖能力的影响.方法 Transwell实验共培养1.0 ×105个MCF-7细胞和1.0×105个成纤维细胞2d,逆转录-聚合酶链反应(RT-PCR)检测成纤维细胞对MCF-7表达WNT10B的影响;三维立体培养2.0×102个MCF-7和5.0×103个成纤维细胞12d,检测MCF-7细胞克隆形成率(CFE).结果 与正常乳腺成纤维细胞(NBFs)共培养2d后,MCF-7细胞中WNT10B的相对表达量为2.1706±0.0646,与癌相关成纤维细胞(CAFs)共培养2d后,MCF-7细胞中WNT10B的相对表达量为3.8642±0.1052,CAFs促进MCF-7细胞表达WNT10B作用更明显(P＜0.05).三维立体共培养实验中,空白对照组中MCF-7细胞的克隆形成率是(6.67±1.89)％,NBFs细胞组中MCF-7细胞克隆形成率是(8.83±2.52)％,与空白对照组比较略有升高,差异无统计学意义(P＞0.05);CAFs细胞组中MCF-7细胞克隆形成率是(37.83±2.02)％,与空白对照组和NBFs组比较均有明显升高,差异有统计学意义(P＜0.05);在CAFs细胞组加入WNT10B单抗后,MCF-7细胞克隆形成率是(11.67±2.56)％,与加入单抗前比较明显降低,差异有统计学意义(P＜0.05).结论 CAFs可以通过上调WNT10B促进乳腺癌细胞增殖.
目的 觀察成纖維細胞對乳腺癌細胞MCF-7錶達WNT10B和增殖能力的影響.方法 Transwell實驗共培養1.0 ×105箇MCF-7細胞和1.0×105箇成纖維細胞2d,逆轉錄-聚閤酶鏈反應(RT-PCR)檢測成纖維細胞對MCF-7錶達WNT10B的影響;三維立體培養2.0×102箇MCF-7和5.0×103箇成纖維細胞12d,檢測MCF-7細胞剋隆形成率(CFE).結果 與正常乳腺成纖維細胞(NBFs)共培養2d後,MCF-7細胞中WNT10B的相對錶達量為2.1706±0.0646,與癌相關成纖維細胞(CAFs)共培養2d後,MCF-7細胞中WNT10B的相對錶達量為3.8642±0.1052,CAFs促進MCF-7細胞錶達WNT10B作用更明顯(P＜0.05).三維立體共培養實驗中,空白對照組中MCF-7細胞的剋隆形成率是(6.67±1.89)％,NBFs細胞組中MCF-7細胞剋隆形成率是(8.83±2.52)％,與空白對照組比較略有升高,差異無統計學意義(P＞0.05);CAFs細胞組中MCF-7細胞剋隆形成率是(37.83±2.02)％,與空白對照組和NBFs組比較均有明顯升高,差異有統計學意義(P＜0.05);在CAFs細胞組加入WNT10B單抗後,MCF-7細胞剋隆形成率是(11.67±2.56)％,與加入單抗前比較明顯降低,差異有統計學意義(P＜0.05).結論 CAFs可以通過上調WNT10B促進乳腺癌細胞增殖.
목적 관찰성섬유세포대유선암세포MCF-7표체WNT10B화증식능력적영향.방법 Transwell실험공배양1.0 ×105개MCF-7세포화1.0×105개성섬유세포2d,역전록-취합매련반응(RT-PCR)검측성섬유세포대MCF-7표체WNT10B적영향;삼유입체배양2.0×102개MCF-7화5.0×103개성섬유세포12d,검측MCF-7세포극륭형성솔(CFE).결과 여정상유선성섬유세포(NBFs)공배양2d후,MCF-7세포중WNT10B적상대표체량위2.1706±0.0646,여암상관성섬유세포(CAFs)공배양2d후,MCF-7세포중WNT10B적상대표체량위3.8642±0.1052,CAFs촉진MCF-7세포표체WNT10B작용경명현(P＜0.05).삼유입체공배양실험중,공백대조조중MCF-7세포적극륭형성솔시(6.67±1.89)％,NBFs세포조중MCF-7세포극륭형성솔시(8.83±2.52)％,여공백대조조비교략유승고,차이무통계학의의(P＞0.05);CAFs세포조중MCF-7세포극륭형성솔시(37.83±2.02)％,여공백대조조화NBFs조비교균유명현승고,차이유통계학의의(P＜0.05);재CAFs세포조가입WNT10B단항후,MCF-7세포극륭형성솔시(11.67±2.56)％,여가입단항전비교명현강저,차이유통계학의의(P＜0.05).결론 CAFs가이통과상조WNT10B촉진유선암세포증식.
Objective To study how fibroblasts can influence breast cancer cells MCF-7 proliferation and the expression of WNT10B.Methods Transwell co-culture of 1.0 × 105 fibroblasts and 1.0 × 105MCF-7 was done for 2 days.Reverse transcription-polymerase chain reaction (RT-PCR) was used to detect the expression of WNT10B in MCF-7 cells.Three-dimensional co-culture 5.0 × 103 fibroblasts and 2.0 ×103 MCF-7 cells were subjected to three-dimensional co-culture for 12 days for assaying the clone formation rate (CFE) of MCF-7 cells.Results Cancer-associated fibroblasts (CAFs) increased significantly the mRNA expression of WNT10B as compared with normal mammary fibroblasts (NBFs) (3.864 2 ± 0.105 2 vs.2.170 6 ±0.064 6,P ＜0.05).CAFs increased significantly the CFE of MCF-7 cells as compared with the controls [(37.83 ± 2.02) ％ vs.(6.67 ± 1.89) ％,P ＜ 0.05],but NBFs did not [(8.83 ± 2.52) ％ vs.(6.67 ± 1.89) ％,P ＞ 0.05],the neutralizing WNT antibody decreased the CFE of MCF-7 cells in CAFs three-dimensional culture [(11.67 ± 2.56) ％ vs.(37.83 ± 2.02) ％,P ＜ 0.05].Conclusion Our results suggest that WNT10B signaling is critical for CAFs-induced growth of human breast cancer cell lines.
