紫杉醇对宫颈癌TC-1细胞表面PD-L1表达的影响

Effect of Paclitaxel on Expression of PD-L1 in Surface of Cervical Cancer TC-1 Cells

目的探讨紫杉醇对宫颈癌TC-1细胞程序性凋亡配体1(PD-L1)表达的影响及其机制。方法 1将细胞分为紫杉醇组及紫杉醇联合蛋白激酶D(PKD)阻断药(G6976),每组设4个浓度梯度,各5个复孔,分别加入对应浓度试剂,MTT法检测紫杉醇对TC-1细胞活力的影响及G6976对紫杉醇IC50值的影响。2将细胞分为0.9%氯化钠溶液组及紫杉醇组,每组5个复孔,分别加入对应浓度试剂,免疫组化法检测紫杉醇对TC-1细胞PD-L1表达的影响。3将细胞分成阴性对照组、G6976组、紫杉醇组、紫杉醇联合G6976组4组,每组5个复孔,分别加入对应浓度试剂,免疫组化法检测紫杉醇、G6976对TC-1细胞PD-L1表达的影响。结果紫杉醇组紫杉醇对TC-1细胞的IC50值为40.0μg·m L-1,紫杉醇联合G6976组紫杉醇对TC-1细胞IC50值为38.9μg·m L-1,两组IC50值差异无统计学意义(P>0.05)。紫杉醇处理TC-1细胞24 h后PD-L1的表达为(88.48±13.44)%,显著高于阴性对照组(39.59±5.99)%(P<0.05)。紫杉醇联合G6976处理TC-1细胞24 h后PD-L1表达为(79.7±4.7)%,显著低于紫杉醇组(96.8±2.5)%(P<0.05)。结论紫杉醇可增加TC-1细胞PD-L1表达,阻断PKD途径可显著抑制PD-L1表达增加的程度,紫杉醇可能是通过PKD途径发挥作用。

Objective To investigate effect of paclitaxel on expression of programmed death ligand-1 （PD-L1） in the surface of cervical cancer TC-1 cells and its mechanism. Methods （1）The cells were divided into two groups ： paclitaxel group, paclitaxel combined with PKD blocker （GO 6976） group.There were 4 concentration gradient and 5 holes for each group, and each hole has its corresponding concentration of drugs.Influence of paclitaxel on TC-1 cell viability and effect of PKD blocker GO 6976 on IC50 value of paclitaxel were evaluated by MTT method.（2）The cells were divided into 0.9% sodium chloride solution （NS） group and paclitaxel group, There were 5 holes of each group.Effect of paclitaxel on PD-L1 expression on the surface of TC-I cells were measured by immunohistochemistry.（3）The cells were divided into 4 groups： NS＋DMSO group, GO 6976 group, paclitaxel group and paclitaxel＋G0 6976 group.There were 5 holes for each group.Effect of paclitaxel and GO 6976 on PD-L1 expression on the surface of TC-1 cells were measured by immunohistochemistry.The expressions of PD-L1 on the surface of cells were measured by immunofluorescence treated with different drugs. Results The IC50 value of paclitaxel was 40.0 μg·m L^-1, in paclitaxel group, and 38.9 μg·m L^-1, in paclitaxel combined with PKD blocker GO 6976 group, without significant difference between the two groups （P〉0.05）.The expression of PD-L1 in the surface of TC-1 cells were significantly higher in paclitaxel group than in negative control group [ （88.48± 13.44）% vs. （39.59±5.99）%, P〈0.05 ] .The expression of PD-L1 in the surface of TC-1 cells was （79.7%±4.7 ）% after treatment with paclitaxel combined with PKD blocker GO 6976 for 24 h, and it was significantly lower than that in paelitaxel group [ （96.8±2.5）%, P〈0.05 ]. Conclusion Paclitaxel promotes the expression of PD-L1 in the surface of TC-1 ceils, which could be significantly inhibited by blocking PKD pathway.Paclitaxel may exert its effect through PKD pathway. KEY WORDS Paclitaxel;Programmed death ligand-1;Protein kinases D;Cancer, cervical