YanQiu Li

Bispecific c-Met/PD-1 CAR-T Cells Have Enhanced Therapeutic Effects on Solid Tumor

Objective To evaluate the killing effect of c-Met CAR-T on tumor cells with different degrees of c-Met expression. It was demonstrated that CAR-T autocrine PD-1 antibody could alleviate immune checkpoint inhibition and enhance the anti-tumor effect of T cells. Methods The specificity and clinical significance of c-Met and PD-L1 expression in various solid tumors were verified by bioinformatics analysis. c-Met specific CAR-T and c-Met specific CAR-T secreted by PD-L1 were synthesized, and c-Met CAR-T and c-Met/PD-1 CAR-T were prepared by constructing lentivirus. Flow cytometry was used to verify the positive rate and cell population of CAR-T, western blot was used to verify the secretion of PD-1 antibody, and cck-8 was used to detect the proliferation of CAR-T in tumor cells with different c-Met expression. LDH and ELISA further evaluated the antitumor effects of c-Met CAR-T and c-Met/PD-1 CAR-T in vitro. Results c-Met and PD-L1 were expressed in pancreatic cancer, ovarian cancer, esophageal cancer, bladder cancer, glioma and other tumors, and were associated with a variety of immune cell infiltration. Tumor cells with high expression of c-Met can strongly stimulate the proliferation of c-Met CAR-T, and c-Met CAR-T has strong cell lysis ability on tumor cells with high expression of c-Met. Autocrine PD-1 antibody can significantly improve the activity of c-Met CAR T cells, tumor lysis ability and cytokine secretion level. Conclusion The antitumor activity of c-Met CAR-T is positively correlated with the expression of c-Met. c-Met CAR-T secreted by PD-1 showed enhanced antitumor function in solid tumor treatment.

Potential serum metabolites and long‐chain noncoding RNA biomarkers for endometrial cancer tissue

AbstractBackgroundEndometrial carcinoma (EC) is one of the most common tumors in the female reproductive system. There are nearly 200 000 new cases every year. It is the third most common gynecological malignant tumor leading to female death. The incidence rate is closely related to lifestyle, and the incidence rate varies in different regions. The incidence rate of EC is ranking the first in the female reproductive system cancer just second only to breast, lung, and colorectal cancer in North America and Europe and the incidence rate of EC is only second, followed by breast cancer and cervical cancer in China.PurposeThe potential metabolic markers of endometrial cancer were screened by liquid chromatograph mass spectrometer (LC‐MS), and the tissues of patients with hysteromyoma and endometrial cancer were sequenced to explore the relationship between the disease and change in the content of long‐chain noncoding RNA (lncRNA).MethodsSerum and tissue samples were collected from patients with endometrial dysplasia, endometrial cancer stage I, and endometrial cancer stage III. The metabolites in all serum samples were extracted, and the metabolites in all samples were detected by LC–MS/MS technology. The Pareto‐scaling method was used for normalization, and the MetaboAnalyst 4.0 software was used for different analyses. The T test between groups showed that p ≤ 0.05 was regarded as the metabolite with a difference. Further, the function of differential metabolites was determined by metabolite function enrichment and co‐expression analysis. Meanwhile, the differentially expressed lncRNA was detected by Illumina second‐generation high‐throughput sequencing technology, and the expression was analyzed by DEGseq software. Different lncRNA were screened according to p < 0.05. LncRNA with significant differences were screened by p < 0.01, q < 0.001, fold change ≥2, and false discovery rate (FDR) ≤0.001.ResultsThrough synthesis of T test, cluster heatmap, and ROC curve analysis, five biomarkers with potential diagnostic ability were obtained, including 2,3‐Pyridinedicarboxylic acid (area under the curve (AUC) = 0.69), Hematommic acid, ethyl ester (AUC = 0.69), Maltitol (AUC = 0.69), 13(S)‐HODE (AUC = 0.88), and D‐Mannitol (AUC = 0.69) had potential diagnostic ability between EC phase I versus EC phase III. At the same time, lncRNA sequencing results showed that when endometrial atypical hyperplasia continued to change, including LINC00511, PVT1, and IQCH‐AS1 (downregulated), and only changed significantly in the endometrial dysplasia group, including MALAT1, CARMN (downregulated) and LINC00648, BISPR, LINC01534, and LINC00930 (upregulated). Moreover, both differential metabolites and differential lncRNA were annotated to the lipid metabolism pathway, suggesting that this pathway played an important role in the occurrence and development of endometrial carcinoma.ConclusionsIt can combine the results of metabolomics and lncRNA sequencing to assist in the early diagnosis of endometrial precancerous lesions and endometrial cancer patients, to enhance the sensitivity and specificity of diagnosis, which has a certain clinical application prospect.

Interleukin‐6 and Hypoxia Synergistically Promote EMT‐Mediated Invasion in Epithelial Ovarian Cancer via the IL‐6/STAT3/HIF‐1α Feedback Loop

Extensive peritoneal spread and capacity for distant metastasis account for the majority of mortality from epithelial ovarian cancer (EOC). Accumulating evidence shows that interleukin‐6 (IL‐6) promotes tumor invasion and migration in EOC, although the molecular mechanisms remain to be fully elucidated. Meanwhile, the hypoxic microenvironment has been recognized to cause metastasis by triggering epithelial–mesenchymal transition (EMT) in several types of cancers. Here, we studied the synergy between IL‐6 and hypoxia in inducing EMT in two EOC cell lines, A2780 cells and SKOV3 cells. Exogenous recombination of IL‐6 and autocrine production of IL‐6 regulated by plasmids both induced EMT phenotype in EOC cells characterized by downregulated E‐cadherin as well as upregulated expression of vimentin and EMT‐related transcription factors. The combined effects of IL‐6 and hypoxia were more significant than those of either one treatment on EMT. Suppression of hypoxia‐inducible factor‐1α (HIF‐1α) before IL‐6 treatment inhibited the EMT phenotype and invasion ability of EOC cells, indicating that HIF‐1α occupies a key position in the regulatory pathway of EMT associated with IL‐6. EMT score was found positively correlated with mRNA levels of IL‐6, signal transducer and activator of transcription 3 (STAT3), and HIF‐1α, respectively, in 489 ovarian samples from The Cancer Genome Atlas dataset. Next, blockade of the abovementioned molecules by chemical inhibitors reversed the alteration in the protein levels of EMT markers induced by either exogenous or endogenous IL‐6. These findings indicate a positive feedback loop between IL‐6 and HIF‐1α, and induce and maintain EMT phenotype through STAT3 signaling, which might provide a novel rationale for prognostic prediction and therapeutic targets in EOC.

ANKRD62 Modulates NF ‐ κB Signaling to Promote Proliferation and Migration in UCEC

ABSTRACT Members of the ankyrin repeat domain (ANKRD) family are involved in multiple cellular functions, including cell cycle regulation, cell adhesion, and signaling, and have been implicated in the pathogenesis of various cancers. However, the function of ANKRD62, a recently identified member of this family, remains largely unexplored. This study investigated the clinical significance and biological function of ANKRD62 in uterine corpus endometrial carcinoma (UCEC). Through gene expression profiling, we identified ANKRD62 upregulation in UCEC tissues compared to paired adjacent normal tissues. Using overexpression and shRNA‐mediated knockdown in cell lines and mouse models, we demonstrated that ANKRD62 is significantly upregulated in UCEC tissues, with expression levels positively correlated with tumor grade and inversely associated with patient survival. In vitro functional assays revealed that ANKRD62 overexpression markedly enhanced UCEC cell proliferation and migration, whereas ANKRD62 knockdown suppressed these malignant phenotypes without significantly affecting apoptosis. Consistent with these findings, in vivo experiments showed that ANKRD62‐overexpressing cells exhibited accelerated tumor growth and increased metastatic potential. Mechanistically, ANKRD62 was found to be associated with NF‐κB pathway modulation, promoting p65 nuclear translocation and phosphorylation, thereby affecting cellular function. In summary, this study provides the first evidence that ANKRD62 acts as a critical oncogenic driver in UCEC progression and unveils its functional association with NF‐κB signaling. These findings highlight the potential of ANKRD62 as a novel therapeutic target and offer new insights into the molecular mechanisms underlying UCEC pathogenesis.

Clinical Studies of Endometrial Cytology and Cervical Methylation Assays in Endometrial Cancer Screening and Fertility-Preservation Evaluation

The current study aims to assess high-risk patients using both liquid-based cytology and cervical methylation testing. The results will be compared with the traditional hysteroscopic pathological findings to determine the sensitivity and specificity of these methods for early detection of endometrial cancer, thereby evaluating their potential application in early screening. Primary Objectives： 1. To evaluate the sensitivity, specificity, and accuracy of endometrial cytology for screening endometrial cancer. 2. To assess the sensitivity, specificity, and accuracy of methylation testing for screening endometrial cancer. 3. To perform further molecular testing on tissue samples obtained from endometrial cytology and cervical methylation tests, aiming to explore early screening-sensitive indicators. Secondary Objectives： 1. To determine the value of endometrial cytology in evaluating the efficacy of fertility-sparing treatments for endometrial cancer. 2. To assess the value of methylation testing in evaluating the efficacy of fertility-sparing treatments for endometrial cancer.