Akt1 induces extracellular matrix invasion and matrix metalloproteinase-2 activity in mouse mammary epithelial cells.
Akt1 induces extracellular matrix invasion and matrix metalloproteinase-2 activity in mouse mammary epithelial cells.
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发表时间：
2001-10
期刊：
影响因子：
11.2
通讯作者：
Bae-Keun Park;Xiao Zeng;Robert I. Glazer
中科院分区：
文献类型：
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作者：
Bae-Keun Park;Xiao Zeng;Robert I. Glazer
蛋白丝氨酸/苏氨酸激酶，Akt 1，在与细胞运动和细胞外基质侵袭相关的信号通路中的作用进行了研究，在永生化小鼠乳腺上皮细胞系，COMMA-1D。COMMA-1D细胞经工程改造以表达禽白血病亚型A受体tv-a，从而允许由复制型禽剪接载体RCAS产生的重组禽白血病病毒感染。用RCAS/v-akt而不是RCAS/Akt 1转导的COMMA-1D/tv-a细胞在软琼脂中形成了锚定非依赖性集落;然而，当在ECM Matrigel上生长时，过表达v-akt或Akt 1的细胞变得高度侵袭性。分泌到培养基中的COMMA-1D/Akt 1或COMMA-1D/v-akt细胞的胞外蛋白酶活性的酶谱显示明胶酶活性升高，通过Western印迹和免疫沉淀酶谱法证实其为基质金属蛋白酶-2（MMP-2;明胶酶A）。MMP抑制剂BB-94可阻断MMP-2活性和与Akt 1和v-akt表达细胞相关的侵袭。蛋白酶体抑制剂，lactacystin，显着增加MMP-2的水平和在对照细胞的侵袭，但不在Akt 1和v-akt表达细胞。这些结果表明，Akt 1诱导的乳腺上皮细胞的侵袭行为与MMP-2的表达增加，这可能是由于抑制MMP-2降解的蛋白酶体途径。
The roles of the protein-serine/threonine kinase, Akt1, in signaling pathways associated with cell motility and extracellular matrix invasion were examined in the immortalized mouse mammary epithelial cell line, COMMA-1D. COMMA-1D cells were engineered to express the avian leukosis subtype A receptor, tv-a, to permit infection by recombinant avian leukosis virus produced by the replication-competent avian splice vector, RCAS. COMMA-1D/tv-a cells transduced with RCAS/v-akt, but not RCAS/Akt1, formed anchorage-independent colonies in soft agar; however, cells overexpressing either v-akt or Akt1 became highly invasive when grown on the ECM, Matrigel. Zymography of extracellular protease activity shed into the medium by COMMA-1D/Akt1 or COMMA-1D/v-akt cells revealed elevated gelatinase activity that was confirmed to be matrix metalloproteinase-2 (MMP-2; gelatinase A) by Western blotting and immunoprecipitation-zymography. The MMP inhibitor, BB-94, blocked MMP-2 activity and invasion associated with Akt1- and v-akt-expressing cells. The proteasome inhibitor, lactacystin, markedly increased MMP-2 levels and invasion in control cells but not in Akt1- and v-akt-expressing cells. These results suggest that the invasive behavior of mammary epithelial cells induced by Akt1 is associated with increased MMP-2 expression that may result from inhibition of MMP-2 degradation by the proteasome pathway.