骨髓激酶X对宫颈癌细胞增殖的影响及可能的作用机制

Effect of bone marrow X-linked kinase on proliferation of human cervical cancer cells and its potential mechanism

目的探讨骨髓激酶X(bone marrow X-linked kinase,BMX)对人宫颈癌细胞增殖的影响及其可能的作用。方法收集2013—2017年本院34例正常宫颈、25例原位宫颈癌和52例浸润性宫颈癌的临床标本,采用免疫组织化学方法检测不同标本中BMX的表达。转染BMX-TALEN重组质粒构建敲低BMX表达宫颈癌细胞HeLa-BMX+/-。宫颈癌细胞HeLa-BMX+/-和HeLa-野生型细胞分别培养于含抑制剂MK-2206和雷帕霉素完全培养基中,并以培养于含有DMSO培养基的细胞为对照组。采用MTT分析测定细胞活力,Western blot检测BMX、Akt、p-Akt、mTOR和p-mTOR表达。结果免疫组织化学法检测结果显示,正常宫颈组织、原位宫颈癌组织及浸润性宫颈癌组织BMX阳性率差异有统计学意义(26.5% vs 68.0% vs 88.5%,χ2=34.804,P<0.001),两两比较差异亦有统计学意义(P<0.05)。成功构建低表达BMX宫颈癌细胞HeLa-BMX+/-。Western blot结果显示,BMX和p-Akt在HeLa-BMX+/-细胞中的表达低于HeLa-野生型细胞(t=6.282,8.117,P<0.001),总Akt表达水平差异无统计学意义(t=2.035,P=0.126)。与对照组比较,经MK-2206和雷帕霉素处理的HeLa-野生型及BMX+/-细胞中p-Akt和p-mTOR的表达均明显受抑制。结论 BMX可能通过PI3K/Akt/mTOR信号通路促进宫颈癌细胞增殖。

Objective To study the role of bone marrow X-linked kinase(BMX) on proliferation of human cervical cancer cells and its potential mechanism. Methods Clinical specimens of 34 cases of normal cervix,25 cases of cervical cancer in situ and 52 cases of invasive cervical cancer were collected from 2013 to 2017. Expression of BMX in different specimens was detected by immunohistochemistry. BMX-TALEN recombinant plasmid was transfected to construct a knockdown BMX-expressing cervical cancer cell line HeLa-BMX+/-. Cervical cancer cells HeLa-BMX+/- and HeLa-wild-type cells were cultured in complete medium containing inhibitor MK-2206 and rapamycin,respectively,and the cells cultured in DMSO medium were used as control group. The cell viability was detected by MTT analysis. The expression of BMX,Akt,p-Akt,mTOR and p-mTOR were detected by Western blot. Results The results of immunohistochemistry showed that the positive rate of BMX in normal cervix,cervical cancer in situ and invasive cervical cancer was statistically significant(26.5% vs 68.0% vs 88.5%,χ2=34.804,P<0.001),and there were statistical differences between the two groups(P<0.005). Low-expression BMX cervical cancer HeLa-BMX cells were successfully constructed. Western blot results showed that the expres-sion of BMX and p-Akt in HeLa-BMX+/- cells were lower than that in HeLa-wild-type cells(t=6.282,8.117,P<0.001),while the expression of total Akt was similar(t=2.035,P=0.126). Compared with the control group,the expression of p-Akt and p-mTOR in HeLa-wild-type and HeLa-BMX+/- cells treated with MK-2206 and rapamycin were inhibited. Conclusion BMX can promote the proliferation of cervical cancer cells,which may be related to the inhibition of PI3K/AKT/mTOR signaling pathway.