Epidermal growth factor receptor (EGFR) mutation testing in plasma cell-free DNA from lung cancer patients is an emerging clinical tool. However, compared with tissue testing, the sensitivity of plasma testing is not yet satisfactory because of the highly fragmented nature of plasma cell-free DNA, low fraction of tumor DNA, and limitations of available detection technologies. We therefore developed a highly sensitive and specific droplet digital PCR method for plasma EGFR mutation (exon19 deletions and L858R) testing. Plasma from 86 EGFR-tyrosine kinase inhibitor-naive lung cancer patients was tested and compared with EGFR mutation status of matched tumor tissues tested by amplification refractory mutation system. By using EGFR mutation-positive cell DNA, we optimized the droplet digital PCR assays to reach 0.04% sensitivity. The plasma testing sensitivity and specificity, compared with the matched tumor tissues tested by amplification refractory mutation system, were 81.82% (95% CI, 59.72%-94.81%) and 98.44% (95% CI, 91.60%-99.96%), respectively, for exon19 deletions, with 94.19% concordance rate (κ = 0.840; 95% CI, 0.704-0.976; P < 0.0001), whereas they were 80.00% (95% CI, 51.91%-95.67%) and 95.77% (95% CI, 88.14%-99.12%), respectively, for L858R, with 93.02% concordance rate (κ = 0.758; 95% CI, 0.571-0.945; P < 0.0001). The reported highly sensitive and specific droplet digital PCR assays for EGFR mutation detection have potential in clinical blood testing.

It is well known that initial consolidation requires de novo gene transcription and protein synthesis in order for memory to become stable. The consolidated memory again becomes labile and temporarily sensitive to disruption when retrieved, requiring a reconsolidation process to become permanent. Although it is well established that glucocorticoid receptors (GR) in the basolateral nucleus of amygdala (BLA) are required for consolidation of fear memory, little is known about their role in reconsolidation of fear memory. In the present study, we first examined the effect of a GR antagonist on postconditioning consolidation of auditory fear memory (AFM). Intra-BLA infusion of the GR antagonist RU486 0 h postconditioning impaired long-term AFM, leaving short-term AFM intact. RU486 had no effect if infusion was performed 6 h postconditioning. We then investigated the effect of the RU486 treatment on postretrieval reconsolidation of AFM. Severe amnesia took place when RU486 was infused into the BLA 0 h postretrieval (reactivation) of AFM, regardless of whether the retrieval was performed 1 day or 10 days postconditioning. RU486 produced no amnesia if the memory retrieval was omitted or if the drug was administered 6 h postretrieval. Treatment with RU486 0 h postretrieval produced no deficit in postretrieval short-term memory but impaired postretrieval long-term memory, and the amnesia exhibited no spontaneous recovery 6 days after retrieval. The present results provide strong evidence that glucocorticoid receptors in the BLA are required for reconsolidation as well as consolidation of AFM.

Homeobox D gene cluster antisense growth‐associated long noncoding RNA (HAGLR) functions as a crucial regulator in the progression and development of human cancers. We analyzed effects of HAGLR, microRNA (miR)‐143‐5p and lysosome‐associated membrane glycoprotein (LAMP)3 on esophageal cancer (EC) and the related mechanisms. Microarray analysis was used to screen out EC‐related genes and the regulation network among HAGLR, miR‐143‐5p, and LAMP3. The regulatory mechanisms of HAGLR and miR‐143‐5p in EC were analyzed following the treatment of miR‐143‐5p mimic, miR‐143‐5p inhibitor, HAGLR vector, or small interfering RNA against HAGLR in EC cells. The expression of N‐cadherin, vimentin, Twist1, Snail1, and E‐cadherin as well as the abilities of cell proliferation, invasion, and migration were measured. The effects of the HAGLR/miR‐143‐5p/LAMP3 axis were determined in vivo by assessing tumor formation in nude mice. The expression of HAGLR and LAMP3 was increased, whereas that of miR‐143‐5p was diminished in EC tissues and cells. HAGLR could competitively bind to miR‐143‐5p, and miR‐143‐5p targeted LAMP3. Down‐regulated HAGLR or up‐regulated miR‐143‐5p increased E‐cadherin expression and significantly diminished expression of LAMP3, N‐cadherin, vimentin, Twist1, and Snail1. Moreover, down‐regulated HAGLR inhibited cell proliferation, invasion, migration, epithelial‐mesenchymal transition (EMT), and tumor growth. Moreover, down‐regulation of HAGLR inhibited LAMP3 expression by sponging miR‐143‐5p, thereby suppressing the progression of EC. Taken together, our results suggest HAGLR acts as a competing endogenous RNA of miR‐143‐5p to increase the expression of LAMP3, thus promoting EMT, proliferation, invasion, and migration in EC cells.—Yang, C., Shen, S., Zheng, X., Ye, K., Sun, Y., Lu, Y., Ge, H. Long noncoding RNA HAGLR acts as a microRNA‐143‐5p sponge to regulate epithelial‐mesenchymal transition and metastatic potential in esophageal cancer by regulating LAMP3. FASEB J. 33, 10490–10504 (2019). http://www.fasebj.org

MiR-135a-5p could repress HNSCC cells proliferation and enhance apoptosis by directly targeting HOXA10, implying miR-135a-5p's significance on HNSCC treatment.

No abstract