COL8A1 siRNA对肝癌细胞体外增殖、侵袭能力及右旋柠烯药敏性的影响

Effects of COL8A1 siRNA on the proliferation, invasion and drug sensitivity to D-limonence treatment in hepatocarcinoma cells

目的应用RNA干扰技术下调小鼠肝癌细胞株（Hca—F）COL8A1的表达，通过观察癌细胞的体外增殖、侵袭能力，研究调节COL8A1表达对肿瘤细胞药敏性的影响。方法采用RT-PCR、WesternBlot分析干扰前后Hca-F细胞中COL8A1的表达情况。用体外侵袭实验检测干扰前后Hca-F细胞的侵袭能力。用MTT法分析干扰前后Hca—F细胞的增殖情况及其对右旋柠烯（D-limonene）的药敏性。结果Hca-F／RNAi细胞中COL8A1的表达与未给予RNA干扰处理的对照组及RNA干扰对照组相比出现明显下调；该细胞的增殖及穿过基质胶的侵袭能力下降；同时上调该细胞对D-limonene的药敏性。结论小鼠肝癌细胞中COL8A1的表达与肿瘤细胞的增殖、侵袭及对抗肿瘤药物的敏感性密切相关，可能为肿瘤的化学治疗提供新靶点。

Objective To investigate the possible effects of COL8A1 on the proliferation, invasion and drug sensitivity of murine hepatocarcinoma cell line Hca-F, we used an RNA interference （RNAi） approach to silence COL8A1 expression. Methods The expression levels of COLSA1 in Hca- F/siRNA cells were assessed by RT-PCR and Western blot. The inhibitory effect of RNAi on Hca-F cell invasion in vitro was demonstrated by ECM invasion assay. The in vitro proliferative ability and drug sensitivity of COL8Al-deficient cells were determined by MTT assay. Results The expression of COLSA1 was significantly reduced in COL8A1/siRNA ceils after 30h transfection, compared with both the RNAi control and the Hca-F ceils. The reduced COLSA1 expression also attenuated the proliferative, invasive ability, as well as increased drug sensitivity of Hca-F/siRNA ceils. Conclusion Our current results indicate that the expression of COL8A1 functionally mediates tumor cell proliferation, invasion, and drug sensitivity, and is a potential target for therapeutic anti-cancer drugs.