基底刚度对人宫颈癌HeLa细胞增殖及顺铂药物敏感性影响的体外研究

The study on the effect of substrate stiffness change on cell proliferation and cisplatin sensitivity of human cervical carcinoma HeLa cells in vitro

目的探讨基底刚度改变对人宫颈癌细胞HeLa增殖活性及对化疗药物顺铂的药物敏感性的影响。方法将人宫颈癌细胞株HeLa细胞培养于基底刚度分别为0.5、5、25 kPa的水凝胶培养皿及基底刚度为106kPa的普通塑料培养皿中。应用CCK8法分别检测不同基底刚度培养皿中细胞的增殖活性;在不同基底刚度的水凝胶培养皿及普通塑料培养皿中分别培养人宫颈癌HeLa细胞,再分别加入不同浓度的顺铂,24 h后应用CCK8法分别检测不同基底刚度组中人宫颈癌HeLa细胞对化疗药物顺铂的药物毒性反应,并计算不同基底刚度组的顺铂半数抑制浓度(IC50)并进行比较。结果在不同基底刚度的培养皿中,在同一时间点随培养皿基底刚度的增加,HeLa细胞的吸光度值越大。在相同基底刚度下,人宫颈癌HeLa细胞存活率随顺铂的药物浓度增加而下降;在相同的顺铂药物浓度下,人宫颈癌HeLa细胞的存活率随基底刚度的增加而下降。0.5、5、25 kPa水凝胶培养皿及普通塑料培养皿中HeLa细胞顺铂IC50值分别为137.20、97.62、52.87、18.40μmol/L。结论刚度较大的基底更利于人宫颈癌HeLa细胞增殖。较大的基底刚度下人宫颈癌HeLa细胞对顺铂更敏感,即增加基底刚度可提高人宫颈癌HeLa细胞对顺铂的敏感性。

Objective To explore the effect of substrate stiffness change on the proliferative activity and cisplatin sensitivity of cervical cancer HeLa cells. Method Human cervical cancer cell line HeLa was cultured in hydrogel culture dishes with the substrate stiffness of 0.5, 5, 25 kPa, as well as in ordinary plastic culture dishes with the substrate stiffness of 106 kPa under same conditions. Cell proliferative activity was detected using the CCK-8;subsequently, cisplatin at different concentrations was added into the hydrogel culture dishes and ordinary plastic culture dish with different substrate stiffness, the toxic reaction in HeLa cells induced by cisplatin in each group after 24 h was detected by CCK-8, besides,the half maximal inhibitive concentration(IC50) of cisplatin in each group was calculated and compared. Result In the culture dishes with differing substrate stiffness, notable increase was observed in HeLa cells assessed by optical density(OD) among all groups at the same time points after culture as the substrate stiffness rose. Additionally, the survival rate of HeLa cells showed a decreasing trend as the cisplatin concentration increased with the same substrate stiffness. IC50 values of cisplatin were 137.20, 97.62, 52.87, 18.40 μmol/L in 0.5, 5, 25 kPa hydrogel culture dishes and ordinary plastic culture dish, respectively. Conclusion Greater substrate stiffness facilitates the cell proliferation of HeLa cells, and also sensitizes HeLa to cisplatin, in other words, increasing substrate stiffness improves the sensitivity of HeLa cells to cisplatin.