Metabolic Reprogramming in Thyroid Cancer.

Thyroid cancer is a common endocrine malignancy with increasing incidence globally. Although most cases can be treated effectively, some cases are more aggressive and have a higher risk of mortality. Inhibiting RET and BRAF kinases has emerged as a potential therapeutic strategy for the treatment of thyroid cancer, particularly in cases of advanced or aggressive disease. However, the development of resistance mechanisms may limit the efficacy of these kinase inhibitors. Therefore, developing precise strategies to target thyroid cancer cell metabolism and overcome resistance is a critical area of research for advancing thyroid cancer treatment. In the field of cancer therapeutics, researchers have explored combinatorial strategies involving dual metabolic inhibition and metabolic inhibitors in combination with targeted therapy, chemotherapy, and immunotherapy to overcome the challenge of metabolic plasticity. This review highlights the need for new therapeutic approaches for thyroid cancer and discusses promising metabolic inhibitors targeting thyroid cancer. It also discusses the challenges posed by metabolic plasticity in the development of effective strategies for targeting cancer cell metabolism and explores the potential advantages of combined metabolic targeting.

[Advances in the targeted treatment of non-small cell lung cancer]. / Fortschritte in der zielgerichteten Behandlung des nicht-kleinzelligen Lungenkarzinoms.

Non-small cell lung cancer (NSCLC) paradigmatically shows the potential of personalized and therefore precise cancer treatment. For around one third of the patients, predominantly suffering from adenocarcinoma, targetable driver mutations could be characterized in the meantime. Targeted therapies, mostly with kinase inhibitors, achieve impressive advances in the prolongation of overall survival often over many years and excellent quality of life in patients with advanced NSCLC. Targeted treatment is also increasingly evaluated as adjuvant or neoadjuvant treatment in early inoperable stages of NSCLC. An absolute prerequisite for the use of personalized treatment is upfront broad molecular diagnostics before the decision on first line treatment. The limitations of personalized treatment are the so far unavoidable development of resistance mutations and increasing clonal heterogeneity during the course of the treatment. Approaches to further improve treatment results comprise the development of next-generation inhibitors, the combination of targeted substances, also with chemotherapy and the use of new immunoconjugates.

Research advances in second-line therapies for hepatocellular carcinoma after resistance to targeted therapy combined with immunotherapy / 临床肝胆病杂志

In recent years， clinical studies on targeted therapy and immunotherapy for advanced hepatocellular carcinoma used alone or in combination have provided abundant evidence on efficacy and safety for the selection of first-line therapies. However， no consensus has been reached on the selection of second-line therapies in various clinical guidelines for hepatocellular carcinoma， which is caused by the fact that existing evidence is limited to the options after failure of sorafenib and that there is still a lack of high-level evidence for new first-line therapies such as second-line therapies after resistance to targeted therapy and immunotherapy for hepatocellular carcinoma. This article reviews the results of current clinical trials and summarizes the studies on second-line therapies for hepatocellular carcinoma after resistance to first-line targeted therapy and immunotherapy for hepatocellular carcinoma based on the different mechanisms of action of drugs， as well as the research advances in recent years. For hepatocellular carcinoma patients with resistance to first-line targeted therapy and immunotherapy， targeted combination therapy and dual-immune therapy are expected to improve treatment outcome and survival， and more prospective clinical studies are needed in the future to provide effective and safe treatment regimens for hepatocellular carcinoma patients with resistance to targeted therapy and immunotherapy.

Research progress in reversing multidrug resistance in colorectal cancer / 国际肿瘤学杂志

Multidrug resistance impacts negatively upon the curative effect of chemotherapy and prognosis in colorectal cancer. There is a growing body of studies trying to develop drugs to overcome multidrug resistance against its common targets, including ATP-binding cassette proteins, metabolic enzymes, apoptotic genes, signaling pathways and genetic material, among which P-glycoprotein inhibitors and drugs disrupting DNA are deeply developed. Developing new inhibitors or combining existing inhibitors with conventional treatment are hopeful ways to overcome multidrug resistance in colorectal cancer.

Expression of p21-activated kinase 2 in laryngeal squamous cell carcinoma and its correlation with clinicopathological characteristics and chemosensitivity / 肿瘤研究与临床

Objective:To investigate the expression of p21-activated kinase 2 (PAK2) in laryngeal squamous cell carcinoma and its relationship with the clinicopathological characteristics and chemosensitivity of patients.Methods:Transcriptome sequencing (RNA-seq) data for laryngeal squamous cell carcinoma were downloaded from the Cancer Genome Atlas (TCGA) database, and 123 patients were included in the study (12 cases had cancer tissues and normal tissues data, and the remaining 111 only had cancer tissues data). Differential expression of PAK2 in cancer and para-cancer tissues was analyzed by using R software, and the potential function of PAK2 in laryngeal squamous cell carcinoma was investigated by using the Kyoto Encyclopedia of Genes and Genomes (KEGG) database signaling pathway enrichment. A total of 34 patients with primary laryngeal squamous cell carcinoma tissues and corresponding para-carcinoma 34 tissue specimens who underwent surgical resection were retrospectively selected from Chaoyang Central Hospital between April 2016 and June 2021, and 20 cases of normal laryngeal mucosa tissues were selected as the controls. Immunohistochemistry was used to detect the expression of PAK2 in various tissues, and its correlation with clinicopathological factors was analyzed. A total of 35 supraglottic primary laryngeal squamous cell carcinoma patients were retrospectively collected before induction chemotherapy during the same period, including 20 patients sensitive to chemotherapy and 15 patients resistant to chemotherapy. Real-time fluorescence quantitative polymerase chain reaction (qRT-PCR) was used to detect the relative expression level of PAK2 mRNA in cancer tissues.Results:Analysis of TCGA database data showed that PAK2 expression was increased in cancer tissues compared with that in para-cancer tissues ( P = 0.012); KEGG database signaling pathways showed that the high expression of PAK2 in laryngeal squamous cell carcinoma was related to signal transduction pathways, cell cycle, and cancer. Immunohistochemistry showed that the proportion of PAK2 positive in 34 cases of laryngeal squamous cell carcinoma tissues was higher than that in adjacent tissues and normal tissues [58.82% (20/34) vs. 0.03% (1/34), 0 (0/20), all P < 0.001]. There were statistically significant differences in the proportion of PAK2 positive patients stratified with different degrees of differentiation [high differentiation vs. low or middle differentiation: 33.33% (6/18)vs. 87.50% (14/16)], lymph node metastasis [presence vs. absence: 90.91% (10/11) vs. 43.48% (10/23)], TNM staging [stage Ⅲ-Ⅳ vs. stage Ⅰ-Ⅱ: 82.35% (14/17) vs. 35.29% (6/17)] (all P < 0.05), and PAK2 positive patients were not associated with clinical type, tumor size, smoking history, drinking history, and age (all P > 0.05). qRT-PCR showed that the relative expression level of PAK2 mRNA in the chemotherapy-resistant group was higher than that in the chemotherapy-sensitive group (3.89±0.12 vs. 0.78±0.23, P < 0.001). Conclusions:The expression level of PAK2 in laryngeal squamous cell carcinoma tissues is increased, and the high expression of PAK2 is closely related to the malignant clinical characteristics of patients with laryngeal squamous cell carcinoma. The high expression of PAK2 may indicate the insensitivity to traditional chemotherapy regimens, and PAK2 may be a potential gene that targets and regulates the chemosensitivity of laryngeal squamous cell carcinoma.

Research progress on the role of non-coding RNA in chemotherapy resistance of osteosarcoma / 中华骨科杂志

Osteosarcoma is the most common primary malignant bone tumor in children and adolescents. With the emergence of chemotherapy resistance in recent years, the survival rate of osteosarcoma patients has reached a bottleneck. Therefore, exploration of its chemoresistance mechanism is one of the popular research directions currently. Non-coding RNA (ncRNA) is a class of RNA without the ability to encode proteins, which is classified into microRNA, long non-coding RNA, and circular RNA based on length and shape. With the development of high-throughput sequencing technology, there is increasing evidence that some non-coding RNAs are abnormally upregulated or downregulated in osteosarcoma cells and affect the response of osteosarcoma to four commonly used chemotherapeutic drugs (methotrexate, doxorubicin, cisplatin and ifosfamide) through mechanisms such as regulation of apoptosis, cell cycle, signaling pathways, intracellular drug concentration, and cellular autophagy. Therefore, thesenon-coding RNAs are expected to be novel targets for osteosarcoma treatment. In this paper, the current studies were searched and reviewed on the above three non-coding RNAs mediating chemoresistance in osteosarcoma, aiming to provide a reference for breaking the bottleneck of survival rate of osteosarcoma patients.

Establishment and preliminary application of organoids in ovarian cancer / 中华妇产科杂志

Objective:To explore the establishment and application of ovarian cancer organoids.Methods:Fresh ovarian tumor tissues, obtaining from patients underwent surgery in the First Affiliated Hospital of Nanjing Medical University between October 2021 and March 2022, were collected, enzymatic degraded, digested, and embedded into matrigel to establish organoids. A total of 32 ovarian cancer samples were collected. Hematoxylin eosin (HE) staining and immunofluorescence (IF) procedure were used to verify the morphological structure of organoids and their expression of molecular markers. 3D cyto-live or dead assay was used to detecte the live or dead cells in organoids. Carboplatin with a concentration ranging from 5 to 80 μmol/L (5, 10, 20, 40, 80 μmol/L) was added to organoids to calculate the 50% inhibitory concentration (IC 50) in different organoids. Results:(1) Organoids from a total of 32 patients were established, of which 18 cases could be passaged stably in the long term in vitro, while 14 could be passaged in the short time. The average amplification time of long-term passage in vitro was over 3 months, and the longest reached 9 months. (2) In HE staining, significant nuclei atypia and local micropapillary structures were observed in organoids. IF staining revealed that ovarian cancer organoids expressed molecular markers similar to primary tumor tissues, such as Pan cytokeratin (Pan-CK), p53, paired box gene 8 (PAX8), and Wilms tumor gene 1 (WT1). (3) In 3D cyto-live or dead assay, a large number of apoptotic cells were observed inside and around the organoids after added carboplatin. The sensitivity to carboplatin varied in 18 organoids could amplify in the long term, with an average IC 50 of (29.5±15.8) μmol/L. Moreover, IC 50 values of 4 organoids derived from patients received neoadjuvant chemotherapy were much higher than the 14 organoids which did not received neoadjuvant chemotherapy [(48.7±11.3) μmol/L vs (24.0±12.1) μmol/L; t=3.429, P=0.022]. Conclusions:Organoids recapitulate ovarian cancers in vitro and could be stably passaged. Organoids derived from patients received neoadjuvant chemotherapy have higher resistance to carboplatin.

[Targeted treatment of non-small cell lung cancer]. / Zielgerichtete Therapie des nichtkleinzelligen Lungenkarzinoms.

Non-small cell lung cancer (NSCLC) has made a remarkable development in recent decades with respect to its perception. In the late 1990s it was the "problem child" as the main cause of cancer with increasing tendencies, especially in women and with a pronounced stigmatization. It is now the role model as a biologically rational targeted treatment based on molecular dependencies of the tumor with a vast improvement of the traditionally poor survival times. Molecular tumor boards have long followed the NSCLC example in the assessment of targeted treatment approaches for other tumor entities. This review article gives an overview of the current possibilities for targeted treatment of NSCLC, which nowadays are applicable for nearly one third of all patients with NSCLC.

PARP inhibitors in ovarian cancer: overcoming resistance with combination strategies.

The use of PARP inhibitors (PARPi) in patients with epithelial ovarian cancer is expanding, with the transition from use in recurrent disease to the first-line setting. This is accompanied with an increasing population of patients who develop acquired PARPi resistance. Coupled with those patients with primary PARPi resistance, there is an urgent need to better understand mechanisms of resistance and identify means to overcome this resistance. Combination therapy offers the potential to overcome innate and acquired resistance, by either working synergistically with PARPi or by restoring homologous recombination deficiency, targeting the homologous recombination repair pathway through an alternate strategy. We discuss mechanisms of PARPi resistance and data on novel combinations which may restore PARPi sensitivity.

Identificação de moduladores da sinalização celular no câncer de cólon e metástases hepáticas como ferramenta para descoberta de alvos terapêuticos / Identification of cell signaling modulators in colon cancer and liver metastases as a tool for discovering therapeutic targets

O câncer de cólon é uma das principais causas de morte por câncer em todo mundo e aproximadamente 50% dos pacientes desenvolvem metástases hepáticas durante o curso da doença. Ainda que a ressecção cirúrgica proporcione sobrevida de 5 anos ao redor de 40%, a maior parte dos pacientes apresenta doença irressecável ao diagnóstico. Neste caso, o cenário é devastador devido à resistência das células tumorais ao tratamento padrão com quimioterapia e/ou anticorpo monoclonal. Apesar dos avanços alcançados pelo rastreamento genômico das metástases, o conhecimento sobre o envolvimento de transcritos e proteínas específicas na atividade de sinalização e recrutamento de populações imunes ainda é limitado nessa doença. Considerando o grande potencial de descoberta e inovação que pode ser alcançado, este projeto propôs o desenvolvimento de uma meta-análise de dados públicos para a exploração de metástases hepáticas e a caracterização proteômica de metástases hepáticas inicialmente irressecáveis em busca de alvos promissores que possam ser explorados em novas opções terapêuticas. As principais etapas metodológicas incluem (1) desenvolvimento de um modelo de integração de dados para a obtenção, processamento e análise de aproximadamente 3.5 mil amostras de RNA-seq, microarray e proteômica de metástases hepáticas inicialmente ressecáveis, tumores primários do cólon e tecidos não-neoplásicos, (2) rastreamento por espectrometria de massas de 30 biópsias pareadas, sendo 15 de metástases hepáticas inicialmente irressecáveis e 15 de tecidos não-neoplásicos adjacentes, (3) investigação de potenciais moduladores da sinalização oncogênica associada à resistência à terapia e (4) exploração dos resultados em um conjunto independente com aproximadamente 138 mil células obtidas por single-cell RNA-seq em busca de associações entre alvos, vias de sinalização e populações imunes. Após a análise de espectrometria de massas e integração de dados da metaanálise, das 46 proteínas diferencialmente reguladas entre metástases hepáticas inicialmente irressecáveis e tecidos não-neoplásicos, TGFB1, CDH2, APOA1, TNFR2, EGFR, MDH2, ITIH2, YWHAZ, PCBP1, TOP2A e ENO1 (todas com aumento de regulação nas metástases) e CXCL14, PLA2G2A e CAV1 (com diminuição de regulação nas metástases) prosseguiram na análise. Dessas, PLA2G2A e CAV1 não foram estatisticamente confirmadas após comparação com cinco estudos independentes. Também foi identificada a ativação nas metástases de importantes vias como JAK-STAT (z-score=3,18), VEGF (z-score=2,67), WNT (z-score=3,22), PI3K (z-score=3,99), MAPK (z-score=2,85), EGFR (z-score=2,49) e NFkB (zscore=4,11). Em seguida observamos pela análise de single-cell RNA-seq que a presença de TGFB1, TNFR2 e EGFR estava associada a maior polarização de neutrófilos e células Tregs em metástases hepáticas, sugerindo uma participação desses alvos no recrutamento dessas populações imunes ou em conjunto durante o desenvolvimento, estabelecimento dessas células no fígado e progressão da doença. Tanto TGFB1 (p=0,01) quanto TNFR2 (p=0,048) foram associados a pior sobrevida global em pacientes metastáticos e apresentaram AUC=0,95 na separação entre câncer primário no cólon e metástases no fígado. Especificamente sobre a comparação entre metástases inicialmente ressecáveis e irressecáveis, apesar de termos identificado alguns alvos com mais de 10-fold de diferença (YWHAZ, AHNAK, HNRNPH1, PCBP1 e TGFB1) apenas TGFB1 prosseguiu em todas as análises, porém, esses resultados sugerem a exploração posterior desses candidatos em novos estudos. Quando buscamos por alvos relacionados à resistência à terapia, nossa estratégia selecionou 32 proteínas diferencialmente reguladas nas metástases resistentes, com envolvimento de CDH2 (p=0,002), CXCL14 (p=0,05), SERPINA1 (p=0,012) e LRGR (p=0,049) com pior sobrevida global em pacientes metastáticos. Nesses tumores também foi observada uma tendência de ativação das vias TGF-ß e TNF-α, já descritas na literatura devido a participação no desenvolvimento e progressão de metástases, além de favorer um perfil de resistência às células tumorais. Por meio de uma análise de deconvolução também observamos maior presença de células Tregs em pacientes resistentes. Essa relação entre Tregs/LGR5 também já foi associada a pior prognóstico em outros tipos de cânceres e suportam uma participação de ambos em um possível mecanismo de resistência desses tumores. Com base nos resultados obtidos, acreditamos oferecer um conjunto de dados inéditos que podem modificar o prognóstico e diagnóstico das metástases hepáticas, bem como contribuir para que os pacientes tenham novas opções terapêuticas com melhora na sobrevida

Colon cancer is one of the leading causes of cancer death worldwide and approximately 50% of patients develop liver metastases during the course of the disease. Although surgical resection provides a 5-year survival rate of around 40%, most patients have unresectable disease at diagnosis. In this case, the scenario is devastating due to the resistance of tumor cells to standard treatment with chemotherapy and/or monoclonal antibody. Despite the advances achieved by genomic tracking of metastases, the involvement of specific transcripts and proteins in the signaling activity and recruitment of immune populations is still limited in this disease. Considering the great potential for discovery and innovation that can be achieved, this project proposed the development of a meta-analysis for the exploration of liver metastases and the proteomic characterization of initially unresectable liver metastases in search of promising targets that can be explored in new therapeutics options. Key methodological steps include (1) development of a data integration model for obtaining, processing and analyzing approximately 3,500 RNA-seq, microarray and proteomic samples from initially resectable liver metastases, primary colon tumors and non-neoplastic tissues, (2) mass spectrometry screening of 30 paired biopsies, 15 from initially unresectable liver metastases and 15 from adjacent non-neoplastic tissues, (3) investigation of potential modulators of oncogenic signaling associated with therapy resistance, and (4) exploration of the results in an independent pool of approximately 138,000 cells obtained by single-cell RNA-seq in search of associations between targets, signaling pathways and immune populations. After mass spectrometry analysis and integration of metaanalysis data, of the 46 differentially regulated proteins between initially unresectable liver metastases and non-neoplastic tissues, TGFB1, CDH2, APOA1, TNFR2, EGFR, MDH2, ITIH2, YWHAZ, PCBP1, TOP2A and ENO1 (all with upregulation in metastases) and CXCL14, PLA2G2A and CAV1 (with downregulation in metastases) continued in the analysis. Of these, PLA2G2A and CAV1 were not statistically confirmed after comparison with five independent studies. We identified activation of important pathways such as JAK-STAT (zscore=3.18), VEGF (z-score=2.67), WNT (z-score=3.22), PI3K (z-score=3.99), MAPK (z- score=2.85), EGFR (z-score=2.49) and NFkB (z-score=4.11). Then we observed by single-cell RNA-seq analysis that the presence of TGFB1, TNFR2 and EGFR was associated with greater polarization of neutrophils and Treg cells in liver metastases, suggesting a participation of these targets in the recruitment of these immune populations or together during development, establishment of these cells in the liver and disease progression. Both TGFB1 (p=0.01) and TNFR2 (p=0.048) were associated with worse overall survival in patients with metastatic disease and had AUC=0.95 separating primary colon cancer from liver metastases. Specifically, regarding the comparison between initially resectable and unresectable metastases, although we identified some targets with more than 10-fold difference (YWHAZ, AHNAK, HNRNPH1, PCBP1 and TGFB1) only TGFB1 continued in all analyses, however, these results suggest the exploration of these candidates in further studies. When looking for targets related to therapy resistance, our strategy selected 32 proteins differentially regulated in resistant metastases, with involvement of CDH2 (p=0.002), CXCL14 (p=0.05), SERPINA1 (p=0.012) and LRGR (p=0.049) with poor overall survival in metastatic patients. In these tumors, a tendency of activation of the TGF-ß and TNF-α pathways was also observed, already described in the literature due to their participation in the development and progression of metastases, in addition to favoring a resistance profile to tumor cells. By means of a deconvolution analysis, we also observed a greater presence of Treg cells in resistant patients. This relationship between Tregs/LGR5 has also been associated with a worse prognosis in other types of cancers and supports the participation of both in a possible mechanism of resistance in these tumors. Based on the results obtained, we believe to offer a set of unpublished data that can modify the prognosis and diagnosis of liver metastases, as well as contribute to patients having new therapeutic options with improved survival

Progress of epidermal growth factor receptor-tyrosine kinase inhibitor targeted therapy in Chinese patients with non-small cell lung cancer / 肿瘤研究与临床

Cell epidermal growth factor receptor (EGFR) mutation is one of the causes of non-small cell lung cancer (NSCLC). The emergence of targeted drugs for EGFR gene mutation provides a new direction for NSCLC treatment. The common EGFR-targeted drugs like the first-generation gefitinib and erlotinib, the second-generation afatinib and the third-generation osimertinib have shown their good efficacies in a number of large international clinical trials. EGFR gene mutation in Chinese NSCLC patients is different from that in European and American NSCLC patients. This paper briefly reviews the characteristics of EGFR gene mutation and the current status and progress of EGFR-targeted drugs in Chinese NSCLC patients to investigate the mutation characteristics of EGFR in Chinese NSCLC patients and the response as well as prognosis of Chinese patients to EGFR-TKI therapy.

Role of thioredoxin reductase 1 in multidrug resistance caused by reactive oxygen species-related cell apoptosis in hepatocellular carcinoma / 临床肝胆病杂志

Objective Drug resistance is the main cause of chemotherapy failure in hepatocellular carcinoma (HCC), and thioredoxin reductase 1 (TXNRD1), as a major influencing factor for reactive oxygen species (ROS) metabolism, has been proven to be associated with the poor prognosis of patients with HCC. This study aims to explore the role of TXNRD1 in the mechanism of multidrug resistance in HCC. Methods BEL/FU cells in BEL-7402 cell line were selected as the multidrug-resistant cell line. The siRNA was used for the intervention of TXNRD1 expression; quantitative real-time PCR and Western blotting were used to measure the expression of TXNRD1; CCK-8 assay and flow cytometry were used to evaluate the effect of TXNRD1 on hepatocyte ROS accumulation, resistance to 5-fluorouracil (5-Fu) and doxorubicin (DOX), and apoptosis in vitro; a xenograft tumor model was established to investigate the effect of auranofin (AUR) on drug resistance in vivo. The two-independent-samples t test was used for comparison of continuous data between two groups. Results As a multidrug-resistant HCC cell line, BEL/Fu showed high mRNA and protein expression levels of TXNRD1 (both P < 0.05). Compared with 5-Fu or DOX treatment alone, the TXNRD1 inhibitor AUR combined with 5-Fu or DOX had had a significant reduction in the number of colony formation ( P < 0.01) and a significant increase in apoptosis ratio ( P < 0.001). The ROS scavenger N-acetylcysteine (NAC) significantly weakened the effect of TXNRD1 knockdown by siRNA on the drug resistance of BEL/Fu cells, and the application of NAC effectively reduced the apoptosis ratio of cells after siRNA interference ( P < 0.001). Animal experiments also confirmed that compared with the nude mice treated with 5-Fu alone, the nude mice treated with 5-Fu and AUR had a significantly lower tumor mass ( P < 0.001) and a significantly smaller tumor volume ( P < 0.001). Conclusion TXNRD1 plays an important role in the drug resistance of HCC, and inhibition of its level in cells can effectively improve drug resistance. As a TXNRD1 inhibitor, AUR has great application prospects in the multimodality therapy for HCC.

The effect of miR-485-5p on cisplatin resistance of colon cancer cells / 中华老年医学杂志

Objective:To investigate the effect of miR-485-5p on cisplatin resistance of colon cancer cells through the PI3K/Akt-PAK1 signaling pathway.Methods:The LoVo/DDP cell lines were constructedand divided into an NC group(without transfection treatment), an miR-485-5p mimics group(transfected with miR-485-5p mimics), an miR-485-5p inhibitors group(transfected with miR-485-5p inhibitors), an IPA-3 group(intervention with IPA-3)and an miR-485-5p mimics+ IPA-3 group(transfection with miR-485-5p mimics andinterventionwith IPA-3), with all given 0, 3, and 5 μmol/L cisplatin treatment.Results:Among the 20 patients, the proportion of negative miR-485-5p detection was 85.0%(17/20), and the proportion of positive detection was 15.0%(3/20); the proportion of negative PAK1 detection was 20.0%(4/20), and the proportion of positive detection was 80.0%(16/20). The expression of miR-485-5p in colon cancer tissue was significantly lower than that in adjacent tissues( P<0.05); the expression of miR-485-5p in the human colon cancer cell lines LoVo, SW620, HCT116, and SW480 was all lower than in normal intestinal mucosal cells( P<0.05); the expression of miR-485-5p in LoVo/DDP was significantly lower than inLoVo( P<0.001). Under the action of 3 μmol/L and 5 μmol/L cisplatin, LoVo/DDP had highercell viability but a lower apoptosis rate than LoVo( P<0.001). The cell survival rate in the miR-485-5p mimics group was significantly lower than that in the miR-485-5p inhibitors group( P<0.001). Compared with the NC mimics group, overexpression of miR-485-5p significantly down-regulated luciferase activity of the wild-type PAK1 reporter gene( P<0.001); P-PI3k, P-Akt and PAK1levels in the miR-485-5p mimics group were significantly lower than in the miR-485-5p inhibitors group( P<0.001); the cell survival rate in the miR-485-5p mimics group, the IPA-3 group and the miR-485-5p mimics+ IPA-3 group was significantly lower than in the NC group( P<0.001)and the cell survival rate in the miR-485-5p mimics+ IPA-3 group was significantly lower than in the miR-485-5p mimics group( P<0.001). Conclusions:Up-regulation of miR-485-5p reverses colon cancer cisplatin resistance through the PI3K/Akt-PAK1signaling pathway, suggesting that overexpression of miR-485-5p or inhibition of the PI3K/Akt-PAK1signaling pathway enhances the therapeutic efficacy of cisplatin in colon cancer.

Effects of autophagy-mediated crizotinib resistance on cancer stem-like cell subsets in anaplastic lymphoma kinase-positive anaplastic large cell lymphoma and its significance / 白血病·淋巴瘤

Objective:To investigate the effects of autophagy-mediated crizotinib resistance on cancer stem-like cell subsets in anaplastic lymphoma kinase (ALK)-positive anaplastic large cell lymphoma (ALK + ALCL). Methods:The preliminary research of our group divided ALK + ALCL Karpas299 cell line into two subgroups: reporter unresponsive (RU) and reporter responsive (RR) cells through the implantation of Sox2 reporter genes, among which the RR cells had the characteristics of stem cells. Fluorescent labeled LC3 overexpressing RR and RU cells (RR-LC3 and RU-LC3) were constructed by lentiviral transfection technique, and the transfection efficiency was verified by using Western blotting and flow cytometry. RU-LC3 and RR-LC3 were treated with crizotinib at different concentrations (0, 250, 500, 1 000 nmol/L). The RED and GEN signals were detected by using double-signal flow cytometry to observe autophagy flux (RED represents the red signal B695 of the next generation of far-red fluorescent protein TagFP635 mKate; GEN represents the green signal from pH-sensitive GFP variant pHluorin B530), and the RED to GEV ratio represents autophagy flux. Real-time quantitative polymerase chain reaction (qRT-PCR) was used to detect autophagy related genes ULK1, WIPI1 and LC3B mRNA expression levels in cells. The effects of different concentrations of crizotinib (250, 500, 1 000 nmol/L) combined with chloroquine (5, 10 μmol/L) on the cell survival were detected by using MTS assay. Results:RU-LC3 and RR-LC3 cells with overexpression of LC3 were successfully constructed. After induction of 250, 500 and 1 000 nmol/L crizotinib, the RED to GEN ratio in RU-LC3 cells was 1.135±0.017, 1.453±0.017 and 1.755±0.021, respectively; the RED to GEN ratio in RR-LC3 cells was 1.193±0.018, 2.116±0.013 and 3.307±0.189, respectively; the RED to GEN ratio in RU-LC3 cells and RR-LC3 cells showed a dose-dependent manner. The RED to GEN ratio in RR-LC3 cells was higher than that in RU-LC3 cells when treated with same concentrations of crizotinib, and the differences were statistically significant (all P < 0.01). The autophagy flux of RR-LC3 cells was larger than that of RU-LC3 cells. When treated without crizotinib, mRNA relative expression levels of ULK1, WIPI1 and LC3B in RR cells were higher than those in RU cells (1.69±0.05 vs.1.01±0.02, t = -1.62, P < 0.01; 1.24±0.04 vs. 1.03±0.05, t = -2.11, P < 0.01; 1.70±0.22 vs. 1.02±0.05, t = -1.74, P = 0.033). In the absence of chloroquine, the half-inhibitory concentration ( IC50) of crizotinib in RR cells was higher than that of RU cells (950 nmol/L vs. 709 nmol/L). After treated with chloroquine, IC50 of RU cells did not change, while IC50 of RR cells was decreased with the increase of chloroquine concentration. Conclusions:Compared with RU cells, autophagy reaction of cancer stem-like RR cells is more rapid and intense, which is considered to be one of the important reasons for their resistance to crizotinib.

Progress in molecular resistance mechanisms of targeted therapy for chronic myeloid leukemia / 白血病·淋巴瘤

Resistance or drug-resistant recurrence of targeted tumor therapy is a complex and multi-factorial process, with the final result of tumor clones that can evade treatment or have relative proliferation advantages under treatment pressure being selectively retained. The BCR::ABL1 fusion gene is the primary molecular abnormality of chronic myeloid leukemia (CML), and the development and application of tyrosine kinase inhibitors (TKI) targeting the BCR::ABL1 fusion protein pioneered the era of small molecule targeted therapeutics. Several TKI have been approved for clinical application or in development. Although most CML patients manifest an excellent response to TKI treatment, there are still some patients with poor primary response or relapse with drug resistance. With the increase in the number of patients with long-term maintenance therapy and the sequential use of multiple TKI, the resistance of TKI has become more complicated. This article introduces the research progress of CML molecular resistance mechanisms in recent years and shares the relevant cutting-edge reports at the 63rd American Society of Hematology Annual Meeting in 2021.

Status and significance of BCR-ABL1 kinase domain mutations in imatinib-resistant chronic myelogenous leukemia and Philadelphia chromosome-positive acute lymphoblastic leukemia patients from Northeast China / 白血病·淋巴瘤

Objective:To explore the characteristics of BCR-ABL1 kinase domain mutations in imatinib-resistant chronic myelogenous leukemia (CML) and Philadelphia chromosome-positive acute lymphoblastic leukemia (Ph + ALL) patients from Northeast China and their impact on prognosis. Methods:The clinical data of 252 CML patients and 49 Ph + ALL patients who were admitted to the First Hospital of Jilin University from January 2013 to October 2018 were retrospectively analyzed. The samples of bone marrow or peripheral blood were collected from patients when imatinib treatment was not effective. Nested polymerase chain reaction (PCR) was used to amplify the BCR-ABL1 kinase domain, and Sequencing Analysis v5.4 software was used to analyze the mutation of BCR-ABL1 kinase domain. Patients were followed up for 6-48 months, and the survival analysis was performed. Results:Among 252 CML patients, the mutations in ABL1 kinase domain were found in 57 patients (22.6%), including 25 patients in the chronic phase, 21 patients in the accelerated phase and 11 patients in the blast crisis; 50 patients had 20 types of single point mutation, and the most common mutation types were E255K (16.0%, 8/50), T315I (14.0%, 7/50), M244V (8.0%, 4/50) and G250E (8.0%, 4/50), which were all concentrated in the P-loop and C-helix domains; 7 patients had double mutations; patients with multiple mutations had the worst prognosis, with a median overall survival (OS) time of 3.2 months. Among 49 Ph + ALL patients, 17 cases (34.7%) were positive for mutations in the BCR-ABL1 kinase domain, 14 patients had 12 types of single point mutation, and 3 patients had multiple mutations; the median OS time of patients with multiple mutations, mutations located in the P-loop and C-helix domains and mutations located in the other domains was 2.0, 8.0 and 18.0 months, and the difference in OS among the three groups was statistically significant ( P < 0.01). Conclusions:Among the imatinib-resistant CML and Ph + ALL patients from Northeast China, point mutations in the P-loop and C-helix domains are most commonly found. Multiple mutations, mutations in the P-loop and C-helix domains are related to the poor prognosis of the patients.

Research status and progress of acute myeloid leukemia stem cells / 中国基层医药

Acute myeloid leukemia is a clonal malignant proliferative disease of myeloid blasts of the hematopoietic system. The leukemia cell population is composed of cells of different stages. Acute myeloid leukemia stem cells are the cells that may cause diseases in immunodeficient animals and can regenerate themselves through continuous transplantation, which causes leukemia. Since more than 96% of leukemia stem cells are in the G0 stage, they may escape the effects of chemical drug stabbing, leading to drug resistance and recurrence of leukemia. Therefore, the key to the treatment of acute myeloid leukemia has always been how to remove leukemia stem cells without damaging hematopoietic stem cells. This review paper addresses the new developments in the immunophenotype of leukemia stem cells and leukemia stem cells-targeting therapy for acute myeloid leukemia.

Ibrutinib in treatment of steroid-resistant chronic graft-versus-host disease: report of 3 cases and review of literature / 白血病·淋巴瘤

Objective:To investigate the clinical characteristics of steroid-resistant chronic graft-versus-host disease (cGVHD) patients and the therapeutic effect of ibrutinib.Methods:The clinical data of 3 steroid-resistant cGVHD patients treated with ibrutinib after allogeneic hematopoietic stem cell transplantation in the First Affiliated Hospital of Soochow University from December 2017 to March 2018 were retrospectively analyzed, and the literature was reviewed.Results:All 3 patients had different degrees of skin and oral cGVHD. One patient's oral symptom improved after the application of prednisone, and the skin symptom was better after oral ibrutinib; one patient's oral symptom improved after oral ibrutinib, but skin symptom did not improve significantly; one patient's skin symptom did not improve significantly. None of the 3 patients presented with adverse reactions such as hemorrhage, infection and cytopenia.Conclusions:Ibrutinib has a certain effect on the improvement of symptoms in steroid-resistant cGVHD patients.

Progress of γ-synuclein in tumor proliferation, invasion, migration and drug resistance / 肿瘤研究与临床

γ-synuclein (SNCG) has been extensively studied for its specific overexpression in various malignant neoplasms. Recently, SNCG has been found to be involved in multiple signaling pathways, including estrogen, AKT-mTOR, mitogen-activated protein kinase (MAPK) and microtubule regulation. SNCG has also been found to be related to the proliferation, invasion, migration and chemotherapy drug resistance of neoplasms. Therefore, SNCG is expected to be the key target of anti-tumor and improving the sensitivity of tumor cells to chemotherapeutic drugs.

Progress of Fibulin protein family in tumor development, progression and chemotherapy resistance / 肿瘤研究与临床

Fibulin protein family is a class of extracellular matrix (ECM) proteins that are widely expressed in various tissues and plays an important role in the formation and stabilization of ECM. More and more studies have found that Fibulin protein family can play a role in tumor suppression or tumor promotion in different tumor tissues, which affects tumor proliferation, invasion and metastasis and is closely related to tumor chemotherapy resistance. Furthermore, Fibulin protein family is expected to be the target of inhibiting and reversing tumor chemotherapy resistance. This article reviews the role, target and molecular mechanism of Fibulin protein family in tumor development, progression and chemotherapy resistance, aiming to find a new research direction for tumor chemotherapy resistance.