骨髓基质抗原蛋白2对甲状腺乳头状癌细胞转移活性的影响

Effect of bone marrow stromal cell antigen 2 on metastatic activity of papillary thyroid cancer cells

目的探究骨髓基质抗原蛋白2(BST2)调节磷脂酰肌醇3激酶/丝氨酸-苏氨酸激酶(PI3K/Akt)信号通路对甲状腺乳头状癌细胞增殖、转移和凋亡的影响.方法选取2022年8月至2023年11月在山西省肿瘤医院诊治的11例甲状腺乳头状癌患者[(65±8)岁]中收集癌组织和癌旁组织.将甲状腺乳头状癌TPC-1细胞,通过随机数表法随机分为4组:阴性对照组,siBST2组,PI3K/Akt信号通路抑制剂组、pcDNA BST2组.免疫组化分析甲状腺乳头状癌组织和癌旁组织BST2的表达水平;CCK-8实验分析各组TPC-1细胞的增殖能力;细胞划痕实验分析各组TPC-1细胞的迁移能力;Transwell实验分析各组TPC-1细胞的侵袭能力;流式细胞术分析各组TPC-1细胞凋亡率;蛋白免疫印迹分析TPC-1细胞中BST2和PI3K/AKT通路蛋白的表达.两组间比较采用独立样本t检验.结果甲状腺乳头状癌组织中BST2表达高于癌旁组织(0.17±0.03比0.83±0.05,t=12.905,P<0.05);siBST2组TPC-1细胞的BST、Akt、PI3K蛋白表达低于阴性对照组(0.60±0.02比0.13±0.02,t=11.367,P<0.05;0.59±0.05比0.11±0.03,t=9.235,P<0.05;0.51±0.06比0.15±0.03,t=15.325,P<0.05);pcDNA BST2组的TPC-1细胞的BST、Akt、PI3K蛋白表达高于阴性对照组(0.60±0.02比0.88±0.55,t=11.911,P<0.05;0.59±0.05比0.81±0.03,t=16.255,P<0.05;0.51±0.06比0.82±0.03,t=13.271,P<0.05);siBST2组、PI3K/Akt信号通路抑制剂组的TPC-1细胞的增殖、迁移、侵袭能力低于阴性对照组(85.12±8.12比153.28±9.88,t=12.766,P<0.05;81.62±9.33比153.28±9.88,t=11.698,P<0.05);siBST2组、PI3K/Akt信号通路抑制剂组的TPC-1细胞的凋亡率高于阴性对照组(13.05±1.16比6.70±0.12,t=9.081,P<0.05;14.36±0.98比6.70±0.12,t=11.661,P<0.05);pcDNA BST2组的TPC-1细胞的增殖、迁移、侵袭能力高于阴性对照组(209.78±15.92比153.28±9.88,t=12.568,P<0.05);pcDNA BST2组的TPC-1细胞凋亡率低于阴性对照组(3.56±0.05比6.70±0.12,t=12.562,P<0.05).结论下调BST2或抑制PI3K/Akt信号通路能够抑制甲状腺乳头状癌TPC-1细胞的增殖、迁移和侵袭能力并促进TPC-1细胞凋亡.

Objective To explore the effect of marrow stromal antigen protein 2(BST2)on prolif-eration,metastasis and apoptosis papillary thyroid cancer cells by regulating phosphatidyl inositol 3 kinase/serine and threonine kinase(PI3K/Akt)signal pathway.Methods Cancer tissues and adjacent tissues were collected from 11 patients(65±8 years old)with papillary thyroid carcinoma diagnosed and treated in Shanxi Cancer Hospital from August 2022 to November 2023.Thyroid papillary carcinoma TPC-1 cells were randomly divided into 4 groups by the random number table method:negative control group,si BST2 group,PI3K/Akt signaling pathway inhibitor group,pcDNA BST2 group.Immunohistochemistry was used to analyze the expression level of BST2 in papillary thyroid carcinoma tissues and adjacent tissues.The pro-liferation ability of TPC-1 cells in each group was analyzed by CCK-8 assay.The migration ability of TPC-1 cells in each group was analyzed by cell scratch test.Transwell assay was used to analyze the invasion ability of TPC-1 cells in each group.The apoptosis rate of TPC-1 cells in each group was analyzed by flow cytome-try.The expression of BST2 and PI3K/Akt pathway proteins in TPC-1 cells were analyzed by Western blotting.Independent sample t test was used for comparison between the two groups.Results The expres-sion of BST2 in papillary thyroid carcinoma tissues was higher than that in adjacent tissues(0.17±0.03 vs.0.83±0.05,t=12.905,P<0.05).The expression of BST,Akt,PI3K proteins in TPC-1 cells of siBST2 group was lower than in the negative control group(0.60±0.02 vs.0.13±0.02,t=11.367,P<0.05;0.59±0.05 vs.0.11±0.03,t=9.235,P<0.05;0.51±0.06 vs.0.15±0.03,t=15.325,P<0.05).The protein expression of BST,Akt and PI3K in TPC-1 cells of pcDNA BST2 group was higher than that of negative control group(0.60±0.02 vs.0.88±0.55,t=11.911,P<0.05;0.59±0.05 vs.0.81±0.03,t=16.255,P<0.05;0.51±0.06 vs.0.82±0.03,t=13.271,P<0.05).Proliferation,migration and invasion ability of TPC-1 cells in siBST2 group and PI3K/Akt signal pathway inhibitors group was lower lower than in the negative control group(85.12±8.12 vs.153.28±9.88,t=12.766,P<0.05;81.62±9.33 vs.153.28±9.88,t=11.698,P<0.05),the apoptosis rate of TPC-1 cells in the siBST2 group and the PI3K/Akt signaling pathway inhibitor group was higher than that in the negative con-trol group(13.05±1.16 vs.6.70±0.12,t=9.081,P<0.05;14.36±0.98 vs.6.70±0.12,t=11.661,P<0.05).The proliferation,migration and invasion abilities of TPC-1 cells in the pcDNA BST2 group were higher than those in the negative control group(209.78±15.92 vs.153.28±9.88,t=12.568,P<0.05).The apoptosis rate of TPC-1 cells in pcDNA BST2 groups was lower than in the nega-tive control group(3.56±0.05 vs.6.70±0.12,t=12.562,P<0.05).Conclusion Down-regulating BST2 or inhibiting PI3K/Akt signaling pathway can inhibit proliferation,migration and invasion ability of TPC-1 cells and promote apoptosis of TPC-1 cells.