Transmembrane member 16 A(TMEM16 A) is involved in many physiological functions, such as epithelial secretion, sensory conduction, nociception, control of neuronal excitability, and regulation of smooth muscle contrac...Transmembrane member 16 A(TMEM16 A) is involved in many physiological functions, such as epithelial secretion, sensory conduction, nociception, control of neuronal excitability, and regulation of smooth muscle contraction, and may be important in peripheral pain transmission. To explore the role of TMEM16 A in the persistent hyperalgesia that results from chronic constriction injury-induced neuropathic pain, a rat model of the condition was established by ligating the left sciatic nerve. A TMEM16 A selective antagonist(10 μg T16 Ainh-A01) was intrathecally injected at L5–6. For measurement of thermal hyperalgesia, the drug was administered once at 14 days and thermal withdrawal latency was recorded with an analgesia meter. For measurement of other indexes, the drug was administered at 12 days,once every 6 hours, totally five times. The measurements were performed at 14 days. Western blot assay was conducted to analyze TMEM16 A expression in the L4–6 dorsal root ganglion. Immunofluorescence staining was used to detect the immunoreactivity of TMEM16 A in the L4–6 dorsal root ganglion on the injured side. Patch clamp was used to detect electrophysiological changes in the neurons in the L4–6 dorsal root ganglion. Our results demonstrated that thermal withdrawal latency was shortened in the model rats compared with control rats.Additionally, TMEM16 A expression and the number of TMEM16 A positive cells in the L4–6 dorsal root ganglion were higher in the model rats, which induced excitation of the neurons in the L4–6 dorsal root ganglion. These findings were inhibited by T16 Ainh-A01 and confirm that TMEM16 A plays a key role in persistent chronic constriction injury-induced hyperalgesia. Thus, inhibiting TMEM16 A might be a novel pharmacological intervention for neuropathic pain. All experimental protocols were approved by the Animal Ethics Committee at the First Affiliated Hospital of Shihezi University School of Medicine, China(approval No. A2017-170-01) on February 27, 2017.展开更多
Uncontrolled cell migration is a common feature of tumor metastasis and formation. Understanding the molecular targets critically involved in cell migration process can lead to the development of potentially novel the...Uncontrolled cell migration is a common feature of tumor metastasis and formation. Understanding the molecular targets critically involved in cell migration process can lead to the development of potentially novel therapeutic strategies for controlling invasion of tumor cells. In this study, we showed that calcium-activated chloride channel ANO 1/TMEM16A played an important role in cell migration and inhibition of ANO1 channel function suppressed the migration of tumor epithelial cells. Silencing ANO 1 by small hairpin RNA （shRNA） resulted in suppression of cell migration and invasiveness in cancer cell lines. In addition, pharmacological inhibition of ANO1 by the channel specific inhibitor T16Ain-A01 significantly slowed down the migration and invasion of tumor epithelial cells in a dose-dependent manner. Taken together, our findings have demonstrated that calcium-activated chloride channel ANO1 contributes to cell migration, and specific ANO1/TMEM16A inhibitors can be the promising candidate to develop new therapies for cancer metastasis.展开更多
基金supported by the National Natural Science Foundation of China,No.30160026(to JQS)the High Level Talent Research Project of Shihezi University of China,No.RCSX201705(to YW)
文摘Transmembrane member 16 A(TMEM16 A) is involved in many physiological functions, such as epithelial secretion, sensory conduction, nociception, control of neuronal excitability, and regulation of smooth muscle contraction, and may be important in peripheral pain transmission. To explore the role of TMEM16 A in the persistent hyperalgesia that results from chronic constriction injury-induced neuropathic pain, a rat model of the condition was established by ligating the left sciatic nerve. A TMEM16 A selective antagonist(10 μg T16 Ainh-A01) was intrathecally injected at L5–6. For measurement of thermal hyperalgesia, the drug was administered once at 14 days and thermal withdrawal latency was recorded with an analgesia meter. For measurement of other indexes, the drug was administered at 12 days,once every 6 hours, totally five times. The measurements were performed at 14 days. Western blot assay was conducted to analyze TMEM16 A expression in the L4–6 dorsal root ganglion. Immunofluorescence staining was used to detect the immunoreactivity of TMEM16 A in the L4–6 dorsal root ganglion on the injured side. Patch clamp was used to detect electrophysiological changes in the neurons in the L4–6 dorsal root ganglion. Our results demonstrated that thermal withdrawal latency was shortened in the model rats compared with control rats.Additionally, TMEM16 A expression and the number of TMEM16 A positive cells in the L4–6 dorsal root ganglion were higher in the model rats, which induced excitation of the neurons in the L4–6 dorsal root ganglion. These findings were inhibited by T16 Ainh-A01 and confirm that TMEM16 A plays a key role in persistent chronic constriction injury-induced hyperalgesia. Thus, inhibiting TMEM16 A might be a novel pharmacological intervention for neuropathic pain. All experimental protocols were approved by the Animal Ethics Committee at the First Affiliated Hospital of Shihezi University School of Medicine, China(approval No. A2017-170-01) on February 27, 2017.
基金The Ministry of Science and Technology of China(Grant No.2014ZX09507003-006-004)
文摘Uncontrolled cell migration is a common feature of tumor metastasis and formation. Understanding the molecular targets critically involved in cell migration process can lead to the development of potentially novel therapeutic strategies for controlling invasion of tumor cells. In this study, we showed that calcium-activated chloride channel ANO 1/TMEM16A played an important role in cell migration and inhibition of ANO1 channel function suppressed the migration of tumor epithelial cells. Silencing ANO 1 by small hairpin RNA （shRNA） resulted in suppression of cell migration and invasiveness in cancer cell lines. In addition, pharmacological inhibition of ANO1 by the channel specific inhibitor T16Ain-A01 significantly slowed down the migration and invasion of tumor epithelial cells in a dose-dependent manner. Taken together, our findings have demonstrated that calcium-activated chloride channel ANO1 contributes to cell migration, and specific ANO1/TMEM16A inhibitors can be the promising candidate to develop new therapies for cancer metastasis.
