PI3K/AKT/mTOR信号通路抑制剂治疗乳腺癌临床应用专家共识

磷脂酰肌醇3-激酶(PI3K)/蛋白激酶B(AKT)/哺乳动物雷帕霉素靶蛋白(mTOR)信号通路(PAM信号通路)在乳腺癌发生发展中发挥重要作用, 与晚期乳腺癌内分泌治疗耐药密切相关, 以PAM信号通路中的关键分子为靶点的抗癌治疗成为了近几年的研究热点。靶向PAM信号通路抑制剂为晚期乳腺癌患者尤其是激素受体阳性人表皮生长因子受体2阴性患者带来明显临床获益。目前, 美国食品和药物管理局批准上市的PAM信号通路抑制剂包括PI3K抑制剂Alpelisib和mTOR抑制剂依维莫司, 同时, 依维莫司也在国内获得批准用于乳腺癌新适应证, Alpelisib有望不久在中国上市。鉴于此, 专家制定了PAM信号通路抑制剂治疗晚期乳腺癌的临床应用专家共识, 系统地介绍了PAM信号通路的机制、PAM信号通路抑制剂的疗效、临床应用、不良事件管理及PIK3CA基因检测建议, 旨在提高中国临床肿瘤医师对PAM信号通路抑制剂的认知, 推进临床决策的精准性, 达到延长患者生存时间的目标。     

PI3K/AKT/mTOR通路调控乳腺癌细胞FASN过表达的分子机制

目的:研究PI3K/AKT/mTOR通路在乳腺癌细胞FASN过表达中的作用,研究抑制该通路能否协同FASN抑制剂发挥靶向抗肿瘤作用。方法:荧光素酶检测法检测mTOR抑制剂rapamycin对乳腺癌细胞FASN转录活性的影响。Western blotting检测PI3K抑制剂LY294002及rapamycin对乳腺癌细胞AKT、pAKT、mTOR通路成员以及FASN的表达影响。分别给予乳腺癌细胞FASN抑制剂cerulenin、rapamycin及两者联合给药,MTT法检测不同干预法对细胞存活的影响。结果:Rapamycin可明显降低乳腺癌细胞FASN启动子活性,其中在FASN过表达的SKBR3细胞中最明显。LY294002作用后pAKT、pmTOR、FASN的表达均明显降低。Ra-pamycin作用后mTOR、pmTOR、p4EBP-1、pp70S6K及FASN的表达均明显下调。Cerulenin可对乳腺癌细胞产生细胞毒作用,在SKBR3中更显著。Rapamycin对乳腺癌细胞的细胞毒作用与FASN的表达无关。Rapam-ycin能够协同cerulenin对乳腺癌细胞产生细胞毒作用,且在SKBR3细胞中更明显,小剂量rapamycin联合小剂量cerulenin即达到较大细胞毒作用。结论:PI3K/AKT/mTOR通路参与调控乳腺癌细胞FASN的过表达,mTOR通路抑制剂联合FASN抑制剂有望成为高侵袭性乳腺癌新的治疗方法。     

GOLM1调控PI3K/AKT/mTOR信号转导通路促进肺腺癌细胞增殖、侵袭和迁移的机制研究

背景与目的：高尔基体膜蛋白1(Golgi membrane protein 1,GOLM1)在肺腺癌中发挥促癌作用,但对肺腺癌细胞增殖、侵袭和迁移的影响及其作用机制尚不明确。探究GOLM1对肺腺癌细胞增殖、侵袭和迁移的影响及其作用机制。方法：选取2019年4月—2021年4月在克拉玛依市中心医院行手术切除的肺腺癌患者的癌组织及相应癌旁组织标本各90例,采用免疫组织化学法检测肺腺癌组织和癌旁组织中GOLM1的表达,并分析肺腺癌组织中GOLM1表达与临床病理学特征的关系。采用蛋白质印迹法（Western blot）检测人肺上皮细胞系BEAS-2B及人肺腺癌H460、A549、PG49、H1299细胞系中GOLM1的表达。取对数生长期的肺腺癌A549细胞,随机分为空白组（细胞未转染）、GOLM1小干扰RNA阴性对照（small interfering RNA negative control,si-NC）组（细胞转染si-NC）、GOLM1小干扰RNA(GOLM1 small interferingRNA,si-GOLM1)组（细胞转染si-GOLM1）、胰岛素样生长因子-1(insulin...     

miR-30a对卵巢癌细胞PTEN/PI3K/AKT/mTOR自噬通路及顺铂耐药性的影响

目的:探究miR-30a对人卵巢癌SKOV3细胞自噬及顺铂耐药性的影响,并初步研究其作用机制。方法:体外培养人卵巢癌SKOV3细胞和人卵巢癌细胞（耐药）细胞（SKOV3/DDP）;将SKOV3/DDP细胞随机分为对照组、miR-30a 阴性对照（NC）组和miR-30a模拟（mimics）组,SKOV3细胞作为正常组。实时荧光定量PCR（RT-qPCR）法检测各组细胞miR-30a表达情况;CCK-8法检测各组细胞顺铂耐药性;Annexin V-FITC/PI法检测各组细胞凋亡情况;蛋白印迹分析法检测各组细胞微管相关蛋白轻链3 I/II（LC3I/II）、p62、磷酸酶和张力蛋白同源物（PTEN）、磷酸化磷脂酰肌醇-3-激酶（p-PI3K）、磷酸化丝氨酸/苏氨酸蛋白激酶B（p-AKT）、磷酸化哺乳动物雷帕霉素靶蛋白（p-mTOR）表达情况;双荧光素酶报告基因检测系统验证miR-30a与PTEN的靶向关系。结果:与对照组和miR-30a NC组相比,miR-30a mimics组SKOV3/DDP细胞miR-30a表达水平、凋亡率、p62、p-PI3K、p-AKT及p-mTOR水平显著升高（P＜0.05）,IC50值、LC3II、PTEN蛋白表达水平及LC3II/LC3I比值显著降低（P＜0.05）;与正常组相比,对照组SKOV3/DDP细胞上述各指标变化趋势完全相反;mirbase数据库预测显示,miR-30a与PTEN mRNA 3' UTR区有结合位点,与PTEN-3' UTR-WT+miR-30a NC组比较,PTEN-3' UTR-WT+miR-30a inhibitor组荧光素酶活性降低（P＜0.05）。结论:miR-30a可能通过靶向抑制PTEN表达,激活PI3K/AKT/mTOR信号通路调控人卵巢癌细胞自噬,降低卵巢癌细胞的顺铂耐药性。     

PI3K/Akt信号转导通路与Basal-like型乳腺癌的分子治疗

磷脂酰肌醇-3激酶/蛋白激酶B (PI3K/Akt)信号转导通路与肿瘤的发生、发展密切相关.研究表明,PI3K/Akt信号通路的活化具有促进乳腺癌细胞增殖及减少细胞凋亡、分化的作用.基底细胞样(Basal-like)乳腺癌是乳腺癌的一种亚型,因其分子表型独特无明确的治疗靶点且恶性程度高、预后差.随着对PI3K/Akt信号转导通路及基底细胞样型乳腺癌发生、发展的研究不断深入,开发应用PI3K/Akt信号通路抑制剂可能为Basal-like型乳腺癌提供有效的分子治疗药物.     

视黄醇结合蛋白7调控Akt/mTOR信号通路对乳腺癌细胞增殖和迁移侵袭的影响

目的 探讨视黄醇结合蛋白7(RBP7)调控蛋白激酶B(Akt)/哺乳动物雷帕霉素靶蛋白(mTOR)信号通路对乳腺癌细胞增殖和迁移侵袭的影响。方法 GEPIA网站用于分析乳腺癌组织的RBP7表达,实时定量PCR(qPCR)用于检测乳腺癌细胞(MCF7、SKBR-3、BT474、T47D、BT549和MDA-MB-231)的RBP7表达。将MDA-MB-231细胞分为pcDNA3.1组(阴性对照)、pcDNA3.1-RBP7组(上调RBP7表达)和pcDNA3.1-RBP7+3BDO组(上调RBP7表达且增强mTOR活性)。qPCR和Western blot用于检测磷酸化Akt(p-Akt)、磷酸化mTOR(p-mTOR)和c-Myc的表达。MTT法、划痕愈合实验和Transwell小室实验检测MDA-MB-231细胞的增殖、迁移和侵袭情况。结果 RBP7在乳腺癌组织和细胞中低表达(P<0.05)。生物信息学分析显示RBP7低表达乳腺癌患者的预后较差。pcDNA3.1-RBP7组p-Akt、p-mTOR和c-Myc的表达低于pcDNA3.1组(P<0.05);而pcDNA3.1...     

PI3K/Akt信号通路在RANKL诱导的MCF-7乳腺癌细胞迁移中的作用

目的：核因子-κB受体活化因子配体（RANKL）能促进表达RANK的上皮癌细胞迁移至骨,与乳腺癌骨转移密切相关。本文探讨磷脂酰肌醇-3-激酶/丝苏氨酸蛋白激酶（phosphoinositide3-kinase/serine/threonine protein kinase PI3K/Akt）信号通路在RANKL诱导的乳腺癌MCF-7细胞迁移中的作用。方法：流式细胞仪检测MCF-7细胞表面RANK蛋白的表达;Transwell法测定RANKL刺激后MCF-7细胞迁移能力的改变;Western-blot检测MCF-7细胞RANKL刺激后p-Akt及Akt的表达。SPSS 16.0软件分析实验数据。结果：MCF-7细胞表达RANK蛋白,RANKL诱导MCF-7细胞迁移能力显著增强,RANKL的圈套受体OPG可阻断RANKL诱导的细胞迁移。RANKL刺激后MCF-7细胞p-Akt表达在1、5分钟时一过性升高,PI3K抑制剂LY294002显著抑制RANKL诱导的细胞迁移。结论：PI3K/Akt信号通路参与RANKL诱导的乳腺癌细胞系MCF-7迁移。     

PI3K/Akt信号通路在RANKL诱导的MCF-7乳腺癌细胞迁移中的作用

目的 核因子-κB受体活化因子配体(RANKL)能促进表达RANK的上皮癌细胞迁移至骨,与乳腺癌骨转移密切相关.本文探讨磷脂酰肌醇-3-激酶/丝苏氨酸蛋白激酶(phosphoinositide3-kinase/serine/threonine protein kinase PI3K/Akt)信号通路在RANKL诱导的乳腺癌MCF-7细胞迁移中的作用.方法 流式细胞仪检测MCF-7细胞表面RANK蛋白的表达;Transwell法测定RANKL刺激后MCF-7细胞迁移能力的改变;Western-blot检测MCF-7细胞RANKL刺激后p-Akt及Akt的表达.SPSS 16.0软件分析实验数据.结果 MCF-7细胞表达RANK 蛋白,RANKL诱导MCF-7细胞迁移能力显著增强,RANKL的圈套受体OPG可阻断RANKL诱导的细胞迁移.RANKL刺激后MCF-7细胞p-Akt表达在1、5分钟时一过性升高,PI3K抑制剂LY294002显著抑制RANKL诱导的细胞迁移.结论 PI3K/Akt信号通路参与RANKL诱导的乳腺癌细胞系MCF-7迁移.     

磷脂酰肌醇-3-激酶/蛋白激酶B/雷帕霉素靶蛋白信号通路与卵巢癌关系的研究进展

卵巢癌是女性生殖系统常见的恶性肿瘤之一,发病率居妇科恶性肿瘤第三位,病死率居妇科恶性肿瘤之首.目前对卵巢癌的标准治疗包括肿瘤细胞减灭术及卡铂和紫杉醇的联合化疗.在卵巢癌的发生过程中,异常信号通路的激活必不可少.磷脂酰肌醇-3-激酶(PI3K)/蛋白激酶B(AKT)/雷帕霉素靶蛋白(MTOR)信号通路在卵巢癌细胞增殖、侵...     

PI3K/AKT信号通路相关蛋白表达与乳腺癌淋巴转移相关性分析

目的:探讨磷脂酰肌醇3-激酶(PI3K)、丝苏氨酸蛋白激酶B(AKT)和血管内皮生长因子受体3(VEGFR-3)在人乳腺癌组织中的表达及其在淋巴转移中的作用。方法:选取60例乳腺癌组织,应用免疫组织化学SP法检测PI3K、AKT和VEGFR-3蛋白的表达,RT-PCR检测PI3K、AKT和VEGFR-3mRNA的表达,并分析其相关性。结果:PI3K在有淋巴结转移组的阳性表达率为78.13%,明显高于无淋巴结转移组53.57%,χ2=4.051 3,P=0.044 1;AKT在有淋巴结转移组的阳性表达率71.88%,明显高于无淋巴结转移组50.00%,χ2=4.029 0,P=0.044 7;VEGFR-3在有淋巴结转移组的阳性表达率为96.89%,明显高于无淋巴结转移组75.00%,χ2=4.435 7,P=0.035 2。60例新鲜标本中,PI3K、AKT和VEGFR-3mRNA在有淋巴结转移的乳腺癌组织中相对表达量分别为0.39±0.17、0.48±0.18和0.42±0.15,明显高于无淋巴结转移组0.31±0.13、0.29±0.23和0.32±0.21,组间比较差异有统计学意义,P值分别为0.047 5、0.000 7和0.036 4。经直线相关分析,PI3K阳性表达组中VEGFR-3的表达为95.00%,PI3K阴性表达组中VEGFR-3表达为70.00%;AKT阳性表达组中VEGFR-3的表达为94.59%,AKT阴性表达组中VEGFR-3的表达为73.91%,在乳腺癌组织中PI3K与VEGFR-3表达呈正相关,r=0.368 5,P=0.003 8;AKT与VEGFR-3表达也呈正相关,r=0.4154,P=0.001。结论:PI3K、AKT和VEGFR-3在乳腺癌组织中的表达呈正相关,PI3K和AKT的表达上调可能促使乳腺癌细胞VEGFR-3过表达,诱导淋巴管生成,促进淋巴转移的发生。     

PI3K/AKT/mTOR pathway activation in primary and corresponding metastatic breast tumors after adjuvant endocrine therapy

Both preclinical and clinical data suggest that activation of the PI3K/AKT/mTOR pathway in response to hormonal therapy results in acquired endocrine therapy resistance. We evaluated differences in activation of the PI3K/AKT/mTOR pathway in estrogen receptor α (ERα) positive primary and corresponding metastatic breast cancer tissues using immunohistochemistry for downstream activated proteins, like phosphorylated mTOR (p‐mTOR), phosphorylated 4E Binding Protein 1 (p‐4EBP1) and phosphorylated p70S6K (p‐p70S6K). For p‐mTOR and p‐4EBP1, the proportion of immunostained tumor cells (0–100%) was scored. Cytoplasmic intensity (0–3) was assessed for p‐p70S6K. The difference between expression of these activated PI3K/AKT/mTOR proteins‐ in primary and metastatic tumor was calculated and tested for an association with adjuvant endocrine therapy. In patients who had received endocrine therapy (N = 34), p‐mTOR expression increased in metastatic tumor lesions compared to the primary tumor (median difference 45%), while in patients who had not received adjuvant endocrine therapy (N = 37), no difference was found. Similar results were observed for p‐4EBP1 and p‐p70S6K expression. In multivariate analyses, adjuvant endocrine therapy was significantly associated with an increase in p‐mTOR (p = 0.01), p‐4EBP1 (p = 0.03) and p‐p70S6K (p = 0.001), indicating that compensatory activation of the PI3K/AKT/mTOR pathway might indeed be a clinically relevant resistance mechanism resulting in acquired endocrine therapy resistance.     

P13K／AKT／mTOR信号通路在乳腺癌Her-2治疗耐药中的作用研究进展

Her-2靶向治疗是Her-2过表达乳腺癌治疗的重要组成部分,但Her-2靶向治疗的耐药严重影响了乳腺癌的治疗。研究证实乳腺癌Her-2靶向治疗出现耐药的过程中有P13K／AKT／mTOR信号通路的激活,因此对P13K／AKT／mTOR信号通路及以P13K／AKT／mTOR信号通路为靶点的药物研究对乳腺癌治疗具有重要意义。     

Targeting the PI3K/AKT/mTOR pathway in estrogen receptor-positive breast cancer

Approximately 70−75% of breast cancers express the estrogen receptor (ER), indicating a level of dependence on estrogen for growth. Endocrine therapy is an important class of target-directed therapy that blocks the growth-promoting effects of estrogen via ER. Although endocrine therapy continues to be the cornerstone of effective treatment of ER-positive (ER+) breast cancer, many patients with advanced ER+ breast cancer encounter de novo or acquired resistance and require more aggressive treatment such as chemotherapy. Novel approaches are needed to augment the benefit of existing endocrine therapies by prolonging time to disease progression, preventing or overcoming resistance, and delaying the use of chemotherapy.     

PI3K/AKT/mTOR通路抑制剂在膀胱癌中的研究进展

磷脂酰肌醇3-激酶(PI3K)/AKT/哺乳动物雷帕霉素靶标(mTOR)通路在人类肿瘤的恶性转化及其随后的生长、增殖和转移中起重要作用。临床前研究表明,PI3K/AKT/mTOR通路在膀胱癌中经常被激活。因此,这一通路被认为是膀胱癌治疗干预的候选通路,针对该通路不同成分的抑制剂正处于临床开发的不同阶段。在这里,重点介绍我们对PI3K/AKT/mTOR通路的最新研究进展,并讨论以该通路为靶点的治疗药物作为膀胱癌治疗药物的发展障碍及发展潜力。     

PI3K/AKT/mTOR信号通路抑制剂在淋巴瘤中的研究进展*

磷脂酰肌醇-3 激酶（phosphatidylinositol 3-kinase,PI 3K）- 蛋白激酶B（protein kinaseB ,PKB ,又称AKT ）- 雷帕霉素靶蛋白（mammalian target of  rapamycin,mTOR）信号通路与细胞的生长、增殖、分化、凋亡、代谢等密切相关,在多种实体肿瘤中已发现该信号通路的异常。近年来,以抑制该通路特定位点的靶向治疗已成为抗肿瘤的研究热点。许多该位点新型抑制剂也已进入淋巴瘤的临床试验中,本文就该通路在淋巴瘤中的活化状态及各个分子靶点抑制剂的研究进展做一综述。     

Multipoint targeting of the PI3K/mTOR pathway in mesothelioma

Background:

Mesothelioma is a notoriously chemotherapy-resistant neoplasm, as is evident in the dismal overall survival for patients with those of asbestos-associated disease. We previously demonstrated co-activation of multiple receptor tyrosine kinases (RTKs), including epidermal growth factor receptor (EGFR), MET, and AXL in mesothelioma cell lines, suggesting that these kinases could serve as novel therapeutic targets. Although clinical trials have not shown activity for EGFR inhibitors in mesothelioma, concurrent inhibition of various activated RTKs has pro-apoptotic and anti-proliferative effects in mesothelioma cell lines. Thus, we hypothesised that a coordinated network of multi-RTK activation contributes to mesothelioma tumorigenesis.

Methods:

Activation of PI3K/AKT/mTOR, Raf/MAPK, and co-activation of RTKs were evaluated in mesotheliomas. Effects of RTK and downstream inhibitors/shRNAs were assessed by measuring mesothelioma cell viability/growth, apoptosis, activation of signalling intermediates, expression of cell-cycle checkpoints, and cell-cycle alterations.

Results:

We demonstrate activation of the PI3K/AKT/p70S6K and RAF/MEK/MAPK pathways in mesothelioma, but not in non-neoplastic mesothelial cells. The AKT activation, but not MAPK activation, was dependent on coordinated activation of RTKs EGFR, MET, and AXL. In addition, PI3K/AKT/mTOR pathway inhibition recapitulated the anti-proliferative effects of concurrent inhibition of EGFR, MET, and AXL. Dual targeting of PI3K/mTOR by BEZ235 or a combination of RAD001 and AKT knockdown had a greater effect on mesothelioma proliferation and viability than inhibition of individual activated RTKs or downstream signalling intermediates. Inhibition of PI3K/AKT was also associated with MDM2-p53 cell-cycle regulation.

Conclusions:

These findings show that PI3K/AKT/mTOR is a crucial survival pathway downstream of multiple activated RTKs in mesothelioma, underscoring that PI3K/mTOR is a compelling target for therapeutic intervention.     

靶向PI3K/AKT/mTOR信号通路治疗急性T淋巴细胞白血病的研究进展

急性T淋巴细胞白血病(T-ALL)是来源于胸腺T细胞祖细胞的具有强侵袭性和异质性的血液系统恶性肿瘤。T-ALL约占儿童急性淋巴细胞白血病(ALL)的15%,在成人ALL中的比例约为25%。强化化疗方案的应用使儿童T-ALL患者预后显著提高,但成人及复发耐药T-ALL患者的预后仍较差。研制新型靶向药物特异性阻断T-ALL细胞内生存及耐药相关的异常激活信号通路近年来被认为是治疗成人及复发难治T-ALL患者的新策略。PI3K/AKT/mTOR通路是T-ALL细胞内异常激活信号通路中具有代表性的一条。目前靶向该通路的多种小分子抑制剂已被成功研制,并在治疗T-ALL的研究中取得良好效果。PI3K/AKT/mTOR通路相关抑制剂较传统化疗药物具有更高的特异性和更低的毒副作用,且诸多研究表明其与低剂量化疗药物或其他靶向药物联合治疗T-ALL能发挥协同效应。本综述将总结近年来在PI3K/AKT/mTOR通路与T-ALL相关领域的研究成果,并对基于靶向该通路治疗T-ALL的研究进展一并阐述。     

Inhibition of squalene epoxidase linking with PI3K/AKT signaling pathway suppresses endometrial cancer

Endometrial cancer (EC) is a common malignant tumor that lacks any therapeutic target and, in many cases, recurrence is the leading ca use of morbidity and mortality in women. Widely known EC has a strongly positive correlation with abnormal lipid metabolism. Squalene epoxidase (SQLE), a crucial enzyme in the cholesterol synthesis pathway regulating lipid metabolic processes has been found to be associated with various cancers in recent years. Here, we focused on studying the role of SQLE in EC. Our study revealed that SQLE expression level was upregulated significantly in EC tissues. In vitro experiments showed that SQLE overexpression significantly promoted the proliferation, and inhibited cell apoptosis of EC cells, whereas SQLE knockdown or use of terbinafine showed the opposite results. Furthermore, we found out that the promotional effect of SQLE on the proliferation of EC cells might be achieved by activating the PI3K/AKT pathway. In vivo, studies confirmed that the knockdown of SQLE or terbinafine can observably inhibit tumor growth in nude mice. These results indicate that SQLE may promote the progression of EC by activating the PI3K/AKT pathway. Moreover, SQLE is a potential target for EC treatment and its inhibitor, terbinafine, has the potential to become a targeted drug for EC treatment.     

Inhibition of PI3K/Akt/mTOR signaling pathway enhances the sensitivity of the SKOV3/DDP ovarian cancer cell line to cisplatin in vitro

The activation of the PI3K/AKT/m TOR pathway plays a key role in ovarian cancer tumorigenesis, progression and chemotherapy resistance. This study aimed to explore the possible mechanism that PI-103, a dual inhibitor of phosphatidylinositide 3-kinase and m TOR, enhances the sensitivity of SKOV3/DDP ovarian cancer cell line to cisplatin chemotherapy. The results showed that PI-103 could significantly increase the sensitivity of SKVO3/DDP cells to cisplatin through inhibiting the activation of PI3K/Akt/m TOR signaling pathway and inducing cell cycle arrest and apoptosis.