酪氨酸激酶受体B在OVCAR3卵巢癌细胞中诱导的失巢凋亡抑制和侵袭

Yu Xiao-Hui, Yang Yi-Xia, Cai Bin, Yan Qin, He Yin-Yan, Wan Xiao-Ping

Department of Obstetrics and Gynecology, Affiliated First People's Hospital, Shanghai Jiao Tong University, Shanghai 200080, China.

Zhonghua Fu Chan Ke Za Zhi. 2008 Sep;43(9):695-9.

OBJECTIVE

To study the relationship between tyrosine kinase receptor B (TrkB) expression and anoikis-suppression and invasion in OVCAR3 ovarian cancer cells.

METHODS

The expression of TrkB mRNA in OVCAR3 ovarian cancer cells under two culture conditions: adhesive cells and cell-spheroids were evaluated by RT-PCR and real-time PCR. The relationship between TrkB expression and anoikis-suppression of OVCAR3 ovarian cancer cells was examined by RNA interference (RNAi) technic, anchorage independent culture and fluorescence-activated cell sorting analysis. The difference in invasion and metastatic ability of OVCAR3 cells under two culture conditions and with or without TrkB silenced by small interfering RNA (siRNA) was investigated by matrigel invasion assay and in vivo studies.

RESULTS

The expression of TrkB mRNA was highest in OVCAR3 ovarian cancer cells, 0.0240 +/- 0.0017, compared with the other three cell lines, 0.0030 +/- 0.0006, 0.0027 +/- 0.0009 and 0.0087 +/- 0.0003 respectively, and the expression in OVCAR3 multicellular spheroids was significantly higher than that in cells under monolayer adhesive culture, 0.0437 +/- 0.0021 versus 0.0240 +/- 0.0017 (P < 0.01). TrkB mediated anoikis-suppression in OVCAR3 ovarian cancer cells. OVCAR3 multicellular spheroids had a higher invasion ability than OVCAR3 cells under monolayer adhesive culture, and the penetrating cells of the two groups were 71.8 +/- 0.8 and 47.7 +/- 0.8 respectively (P < 0.01). The metastatic ability of OVCAR3 cells was attenuated when TrkB was silenced, and the volume of the tumors developed by OVCAR3 adhesive cells and OVCAR3 adhesive cells with TrkB silenced were (16.3 +/- 4.7) mm(3) and (6.0 +/- 1.4) mm(3) respectively (P < 0.01).

CONCLUSION

As an anoikis-suppressor, TrkB may increase the invasion and metastasis of OVCAR3 ovarian cancer cells.

目的

研究酪氨酸激酶受体B（TrkB）表达与OVCAR3卵巢癌细胞失巢凋亡抑制及侵袭之间的关系。

方法

采用RT-PCR和实时PCR评估在两种培养条件下（贴壁细胞和细胞球状体）OVCAR3卵巢癌细胞中TrkB mRNA的表达。通过RNA干扰（RNAi）技术、非锚定依赖培养和荧光激活细胞分选分析，研究TrkB表达与OVCAR3卵巢癌细胞失巢凋亡抑制之间的关系。通过基质胶侵袭试验和体内研究，探讨在两种培养条件下以及用小干扰RNA（siRNA）沉默或未沉默TrkB时OVCAR3细胞侵袭和转移能力的差异。

结果

OVCAR3卵巢癌细胞中TrkB mRNA的表达最高，为0.0240±0.0017，而其他三种细胞系分别为0.0030±0.0006、0.0027±0.0009和0.0087±0.0003；OVCAR3多细胞球状体中的表达显著高于单层贴壁培养的细胞，分别为0.0437±0.0021和0.0240±0.0017（P<0.01）。TrkB介导OVCAR3卵巢癌细胞的失巢凋亡抑制。OVCAR3多细胞球状体比单层贴壁培养的OVCAR3细胞具有更高的侵袭能力，两组的穿膜细胞数分别为71.8±0.8和47.7±0.8（P<0.01）。当TrkB被沉默时，OVCAR3细胞的转移能力减弱，OVCAR3贴壁细胞和沉默TrkB的OVCAR3贴壁细胞形成的肿瘤体积分别为（16.3±4.7）mm³和（6.0±1.4）mm³（P<0.01）。