僵蚕含药血清对肝癌细胞侵袭能力的影响

Influence of Serum Containing Stiff Silkworm [Bombyx Batryticatus] on Invasion Ability of Hepatocarcinoma

目的探讨僵蚕抗肝癌转移的可能作用机制。方法 10只SD大鼠给予630 mg/100 g僵蚕水煎剂溶液灌胃,10只大鼠给予等体积生理盐水灌胃,连续灌胃3天后1h心脏采血,制备大鼠含药血清。将肝癌Hepa1-6细胞分为对照组和实验组,分别给予对照血清和僵蚕含药血清培养。用5-乙炔基-2-脱氧尿苷(Ed U)染色法检测各组Hepa1-6细胞染色阳性率判定增殖情况;用Transwell小室法检测各组细胞侵袭能力;设立空白组,采用RT-PCR法检测Hepa1-6细胞上皮细胞间充质转化标志物E-cadherin、N-cadherin mRNA水平。结果实验组肝癌细胞Hepa1-6 Ed U染色阳性率低于对照组,肝癌细胞侵袭数目亦低于对照组(P<0.01)。与空白组比较,对照组E-cadherin mRNA下调,N-cadherin mRNA上调(P<0.05);与对照组比较,实验组E-cadherin mRNA表达增加,N-cadherin mRNA表达减少(P<0.05)。结论僵蚕含药血清能抑制肝癌细胞Hepa1-6体外的增殖、侵袭,可能通过减弱上皮细胞间充质转化发挥抑制肝癌转移的作用。

Objective To explore the potential mechanism of the resistance of Jiangcan（ stiff silkworm） to liver cancer metastasis. Methods Ten Sprague Dawley rats were given 630 mg/100 g solution of stiff silkworm decoction orally,while another ten rats given equal volume of normal saline by gavage. They were treated for 3 days. One hour after the administration on the 3 rd day,blood from heart was drawn to prepare the rat serum containing medicine.Hepa1-6 cells were divided into a control group and an experimental group,and were respectively cultured by the control serum and serum containing stiff silkworm. 5-ethynyl-2-deoxyuridine（ Ed U） staining was used to detect the positive staining rate of Hepa1-6 cells in each group to determine the proliferation; Transwell chamber assay was used to detect cell invasion ability; a blank group was established,and mRNA expressions of E-cadherin and N-cadherin in Hepa1-6 cells were detected by RT-PCR. Results The positive rate of Hepa1-6 Ed U staining in experimental group was lower than that in the control group,and the hepatocarcinoma invasion number was also lower than that in the control group（ P 0. 01）. Compared with the blank group,E-cadherin mRNA was down-regulated and N-cadherin mRNA were up-regulated in the control group（ P 0. 05）; compared with the control group,the expression of E-cadherin mRNA increased,and the expression of N-cadherin mRNA decreased in the experimental group（ P 0. 05）. Conclusion Stiff silkworm-containing serum can inhibit the proliferation and invasion of Hepa1-6 in vitro,which may inhibit the metastasis of hepatocellular carcinoma by attenuating the epithelial mesenchymal transition.