Basic research of annexin V as a molecular target therapy for gynecologic cancer

近年来，肿瘤细胞增殖和发展的分子机制(浸润性和侵袭性)已被人们所熟知。因此，肿瘤患者的分子靶向治疗正在欧美国家蓬勃发展。一些酪氨酸激酶抑制剂已经作为抗癌药物进行了临床试验。代表丝氨酸/苏氨酸蛋白激酶的蛋白激酶C(PKC)是最重要的蛋白激酶之一，但尚未发现细胞内的内源性抑制物。我们首次从人胎盘中分离和纯化了膜联蛋白V，它是一种依赖于钙离子的磷脂结合蛋白，据报道在体外可以抑制蛋白激酶C的活性。本研究的目的是阐明Annexin V在细胞中的作用，并阐明Annexin V是否能抑制细胞内的肿瘤进展，进而开发应用丝氨酸/苏氨酸蛋白激酶抑制剂抑制肿瘤进展的分子靶向治疗癌症患者。我们构建了附件…在V-稳定表达的人白血病细胞系(HL-60)中更多。虽然Annexin V过表达不会引起细胞形态的改变，但在Annexin V过表达的细胞中，TPA诱导的PKC活化被抑制。我们发现PKCδ是最有可能的针对膜联蛋白V抑制的蛋白激酶C亚型的候选蛋白，由于膜联蛋白V的过度表达不能诱导表型，因此我们通过阐明膜联蛋白V基因的调控来探讨膜联蛋白V的生物学功能。我们的结果提示：(1)蛋白质合成抑制剂(CHX、ANI)可明显诱导Annexin V基因的表达。(2)TPA通过激活PKC和mRNA转录，增强CHX诱导的MCAS细胞Annexin V mRNA的超诱导。(3)ANI通过ERK1/2 MAPK途径，而不是通过p38 MAPK途径，上调Annexin V mRNA的表达。综上所述，这些结果表明Annexin V基因表达的调控在转录因子水平上是负调控的，PKC和ERK1/2通路参与了Annexin V基因表达的诱导。较少

Recently, molecular mechanism of proliferation and progression of carcinoma cells (infiltration and invasion) has been well understood. As a result, molecular target therapy of cancer patient is developing in Europe and the United States. Clinical trials of some tyrosine kinase inhibitors have already carried out as anti -cancer drugs. Although protein kinase C (PKC) representing Ser/Thr protein kinases is one of the most important kinases, no endogenous inhibitor in the cell has not been discovered. We first separated and purified annexin V from human placenta, which was a Ca^<2+>-dependent phospholipid binding protein, and reported to inhibite protein kinase C activity in vitro. The purpose of this study is to elucidate the role of annexin V in the cell and to clarify whether annexin V can inhibit tumor progression in the cell, followed by developing molecular target therapy of cancer patients with Ser/Thr protein kinase inhibitors by inhibiting tumor progression.We constructed annex … More in V-stably expressing human leukemia cell lines (HL-60). Although annexin V overexpression did not induce morphological changes in the cell lines, TPA-induced PKC activation was inhibited in annexin V overexpressing cells. We showed that PKCδ was a most likely, candidate of TPA targeting PKC isoforms that was inhibited by annexin V. Since annexin V overexpress ion did not induce phenotype, we approached biological function of annexin V by the elucidation of annexin V gene regulation. Our results suggest that (i)protein synthesis inhibitors (CHX, ANI) superinduce annexin V mRNA expression. (ii)TPA enhances CHX-induced the superinduction of annexin V mRNA in MCAS cells by activating PKC and mRNA transcription. (iii)ANI superinduces annexin V mRNA expression through the ERK1/2 MAPK pathway, but not through the p38 MAPK pathway. Taken together, these results indicated that the regulation of annexin V gene expression was negatively controlled at the transcription factor level, and that PKC and ERK1/2 pathway was involved in the induction of annexin V gene expression. Less