酪氨酸激酶3对赫赛汀耐药的卵巢癌细胞株SKOV3/H增殖和成瘤能力的影响

Effect of HER-3 on proliferation and tumorigenicity abilities of Herceptin-resistant ovarian cancer cell line SKOV3/H

摘要:

探索erb-b2受体酪氨酸激酶3（HER-3）对卵巢癌耐药细胞株SKOV3/H的体外增殖和体内成瘤能力的影响。方法采用赫赛汀浓度递增法构建SKOV3 耐药细胞株（赫赛汀起始浓度为5 mg/L，经5～8 次浓度递增，每次提高50%）。实时荧光定量聚合酶链反应（qRT-PCR）检测HER-3在15 对非耐药卵巢癌组织和耐药卵巢癌组织中的表达（SKOV3 和SKOV3/H 细胞中的表达）。MTT 实验验证分别转染sh-HER-3 及其对照质粒vector对SKOV3/H细胞增殖能力的影响。将包含sh-HER-3的重组慢病毒质粒及其阴性对照慢病毒空载体质粒vector（均带egfp荧光标签）分别以病毒/细胞数量=15 的比例感染SKOV3/H 细胞，加入2.0μg/ml 的嘌呤霉素筛选，构建稳定转染sh-HER-3 或vector 的SKOV3/H 细胞，将2 株细胞分别注射到裸鼠皮下，复制卵巢癌裸鼠移植瘤模型，观察体内成瘤情况。结果成功构建卵巢癌SKOV3耐赫赛汀细胞株SKOV3/H，且SKOV3/H的群体倍增时间为SKOV3 细胞的1.4～1.9倍。HER-3在耐药卵巢癌组织中的表达水平高于非耐药癌组织（p <0.05），且HER-3 在耐药细胞株SKOV3/H中的表达水平也高于非耐药卵巢癌细胞株SKOV3（p <0.05）。转染sh-HER-3后，SKOV3/H细胞的增殖能力低于阴性对照组（p <0.05）。转染sh-HER-3后，耐药细胞株SKOV3/H的裸鼠体内成瘤能力降低（p <0.05）。结论HER-3参与卵巢癌的赫赛汀耐药，并正向影响耐药细胞株SKOV3/H的增殖和体内成瘤能力。

Abstract:

To explore the role of HER-3 in ovarian cancer and Herceptin resistant ovarian cancer, and the effect of HER-3 on the proliferation abilities in vitro and tumorigenicity abilities in vivo of Herceptin-resistant ovarian cancer cell SKOV3/H. Methods Herceptin concentration was gradually increased to construct resistant cell lines (C0 = 5 mg/L, increased 5 to 8 times, each time increased by 50%). qRT-PCR was used to detect the expression of HER-3 in 15 pairs of ovarian cancer tissues which were non-resistant or resistant to Herceptin, and the expression of HER-3 in SKOV3 and SKOV3/H cells. MTT experiment was used to detect the proliferation of SKOV3/H after infected with sh-HER-3 or vector. KOV3/H cells infected with sh-HER-3 or vector were subcutaneously injected into nude mice to build the ovarian cancer xenografts. Tumor formation was observed in vivo situation. Results Successfully constructed the Herceptin-resistant ovarian cancer cell line SKOV3/H, the Td of SKOV3/H was 1.4 to 1.9 times of SKOV3. The expression levels of HER-3 was significantly higher in Herceptin resistant ovarian cancer tissues (p < 0.05), and HER-3 expression levels were also significantly higher in SKOV3/H cells ( p< 0.05). After transfected with sh-HER-3, theproliferation and tumorigenicity abilities of SKOV3/H were significantly lower than vector group (p < 0.05). The tumorigenicity ability of SKOV3/H in mice was significantly reduced (p < 0.05). Conclusions HER-3 is involved in the Herceptin resistant of ovarian cancer and affects the proliferation and in vivo tumorigenicity of SKOV3/H.