顺铂联合米托蒽醌调节脑胶质瘤U87细胞Sonic Hedgehog信号通路的研究

目的：观察米托蒽醌（Mitoxantrone,MXT）联合顺铂（Cisplatin,CDDP）对脑胶质瘤U87细胞杀伤活性及对Sonic Hedgehog信号通路的影响。方法：应用MTT法检测不同浓度米托蒽醌、顺铂以及两药物联合对U87细胞成活率的影响。显微镜观察细胞的形态变化DiOC6荧光染料对细胞线粒体染色检测其膜电位变化来反映细胞凋亡。RT-PCR法检测顺铂、米托蒽醌及两药联合对U87细胞Gli1和Ptch基因表达的影响。结果：MTT结果显示顺铂、米托蒽醌均可以有效抑制U87细胞的增殖,当米托蒽醌和顺铂浓度≤0.625μg/ml时,两药联合对U87细胞增殖具有协同抑制作用;细胞形态变化及线粒体膜电位结果显示,单药处理可促进U87细胞凋亡,而联合用药可以协同促进U87细胞的凋亡;RT-PCR法检测显示顺铂对Gli1基因的表达有上调作用,而米托蒽醌、米托蒽醌联合顺铂能下调Ptch和Gli1基因的表达。结论：胶质瘤U87细胞在化疗药物米托蒽醌以及两药物联合作用下可影响Sonic Hedgehog信号通路,协同发挥其促凋亡作用,进而增强肿瘤细胞对化疗药物的敏感性。     

卵巢和卵巢肿瘤中Hedgehog 信号通路的表达

目的探讨正常卵巢和卵巢肿瘤中Hedgehog信号通路的表达。方法采用RT—PCR和免疫组织、细胞化学染色检测26例卵巢肿瘤组织、3例卵巢癌细胞株（SKOV3、A2780和COC1）和7例正常卵巢组织中Hedgehog信号通路的信号分子SHH、IHH、PTCH、SMO和GLI1的mRNA和蛋白表达情况;通过实时定量RT—PCR对随机抽取的11例卵巢肿瘤组织和5例正常卵巢组织进行PTCH和GLI1 mRNA水平的比较。结果SHH、IHH、PTCH、SMO和GLI1的mRNA在卵巢肿瘤组织中均有表达;SHH mRNA在正常卵巢组织中不表达,而IHH、PTCH、SMO和GLI1的mRNA在正常卵巢组织中有表达;在卵巢肿瘤组织中,PTCH和GLI1的mRNA表达水平高于正常组织（P〈0．05）。SHH、IHH、PTCH、SMO和GLI1蛋白在卵巢肿瘤组织中均呈阳性表达,在正常卵巢组织中不表达。卵巢癌细胞株SKOV3、A2780和COC1均有IHH、PTCH、SMO和GLI1的mRNA和蛋白的表达;SHH仅在SKOV3和A2780中表达。结论卵巢肿瘤中存在Hedgehog信号通路,在部分卵巢肿瘤中有Hedgehog信号通路过度激活的现象。     

Hedgehog signalling in androgen independent prostate cancer

Objectives

Androgen-deprivation therapy effectively shrinks hormone-naïve prostate cancer, both in the prostate and at sites of distant metastasis. However prolonged androgen deprivation generally results in relapse and androgen-independent tumour growth, which is inevitably fatal. The molecular events that enable prostate cancer cells to proliferate in reduced androgen conditions are poorly understood. Here we investigate the role of Hedgehog signalling in androgen-independent prostate cancer (AIPC).

Methods

Activity of the Hedgehog signalling pathway was analysed in cultured prostate cancer cells, and circulating prostate tumour cells were isolated from blood samples of patients with AIPC.

Results

AIPC cells were derived through prolonged culture in reduced androgen conditions, modelling hormone therapy in patients, and expressed increased levels of Hedgehog signalling proteins. Exposure of cultured AIPC cells to cyclopamine, which inhibits Hedgehog signalling, resulted in inhibition of cancer cell growth. The expression of the Hedgehog receptor PTCH and the highly prostate cancer–specific gene DD3^PCA3 was significantly higher in circulating prostate cancer cells isolated from patients with AIPC compared with samples prepared from normal individuals. There was an association between PTCH and DD3^PCA3 expression and the length of androgen-ablation therapy.

Conclusions

Our data are consistent with reports implicating overactivity of Hedgehog signalling in prostate cancer and suggest that Hedgehog signalling contributes to the androgen-independent growth of prostate cancer cells. As systemic anti-Hedgehog medicines are developed, the Hedgehog pathway will become a potential new therapeutic target in advanced prostate cancer.     

Hedgehog Signaling in Mammary Gland Development and Breast Cancer

The Hedgehog pathway is critical for many developmental processes, including the formation of several epidermal appendages. In the mammary gland strict regulation of the Hedgehog pathway is required for normal development. Alterations in Hedgehog signaling result in defects in both the embryonic and postnatal mammary gland. Activation of Hedgehog signaling either by mutation or misexpression of pathway members can lead to the development and/or progression of cancers in multiple organs. This review addresses the current understanding and controversies of Hedgehog signaling in mammary gland development and its potential role in promoting breast carcinogenesis and cancer progression.     

Forsaken Pharmaceutical: Glasdegib in Acute Myeloid Leukemia and Myeloid Diseases

Advancements in the understanding of the pathogenesis of acute myeloid leukemia (AML) have led to the introduction and approval of a number of novel drugs in AML. Glasdegib, an oral hedgehog pathway inhibitor, was approved in 2018 in combination with low-dose cytarabine for the treatment of newly diagnosed AML in patients unfit for intensive chemotherapy. In this review, we discuss the preclinical rationale for glasdegib, important clinical trials that led to glasdegib’s approval, and future trials of glasdegib in AML and other myeloid diseases. Notably, 2 large randomized, placebo-controlled phase 3 trials (AML BRIGHT 1019) are currently recruiting patients with newly diagnosed AML to evaluate glasdegib in combination with intensive chemotherapy or azacitidine, depending on the patient’s ability to tolerate induction chemotherapy. While glasdegib and low-dose cytarabine have been eclipsed by venetoclax and hypomethylating agent combinations for newly diagnosed AML in the United States, we discuss other areas where glasdegib may still have an opportunity to improve outcomes in this devastating disease.