2021Molecular Genetics & Genomic MedicineOpen access

LncRNA‐H19 gene plays a significant role in regulating glioma cell function

Ping Liu, Xinqiong Huang, Haijun Wu, Guoling Yin, Liangfang Shen

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Abstract

BACKGROUND: Glioma is an aggressive adult primary cancer, and is characterized by low cure rate, poor prognosis, and high recurrence. The present study aimed to investigate the effect of lncRNA-H19 gene silencing on glioma cell function. METHODS: lncRNA-H19 interference vector (LV3-si-H19) and negative control vector (LV3-NC) were stably transfected into U251 and U87-MG cells, respectively. Quantitative real-time PCR (qRT-PCR) was performed to investigate the expression of lncRNA-H19. Cell proliferation capacity was tested by adopting cell counting kit (CCK8), and propidium iodide (PI) was used for cell cycle analysis. Meanwhile, flow cytometry (FCM) method was used to investigate cell apoptosis, cell migration capacity was detected via wound healing and transwell experiments, and sphere-forming ability was examined in serum-free suspension culture. Additionally, glioma animal models were conducted through injecting U251 cells to estimate the effects of lncRNA-H19 on glioma growth in vivo. RESULTS: Knocking down lncRNA-H19 gene could effectively suppress the proliferation of U251 and U87-MG cells. The knockdown of lncRNA-H19 remarkably inhibited the migration and blocked cycle progressions of U251 and U87-MG cells, yet, no obvious changes were observed in cell apoptosis. Besides, inhibiting lncRNA-H19 expression could attenuate sphere-forming function of U251 and U87-MG cells. Additionally, tumor volume and weight were significantly reduced in rats injected with U251 LV-si-H19 cell line compared to untransfected and negative controls, when survival time was obviously prolonged in U251 LV-si-H19 injection groups. CONCLUSION: LncRNA-H19 gene plays a carcinogenic role in glioma progression via enhancing aggressive behavior of glioma cells.

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BACKGROUND: Glioma is an aggressive adult primary cancer, and is characterized by low cure rate, poor prognosis, and high recurrence. The present study aimed to investigate the effect of lncRNA-H19 gene silencing on glioma cell function. METHODS: lncRNA-H19 interference vector (LV3-si-H19) and negative control vector (LV3-NC) were stably transfected into U251 and U87-MG cells, respectively. Quantitative real-time PCR (qRT-PCR) was performed to investigate the expression of lncRNA-H19. Cell proliferation capacity was tested by adopting cell counting kit (CCK8), and propidium iodide (PI) was used for cell cycle analysis. Meanwhile, flow cytometry (FCM) method was used to investigate cell apoptosis, cell migration capacity was detected via wound healing and transwell experiments, and sphere-forming ability was examined in serum-free suspension culture. Additionally, glioma animal models were conducted through injecting U251 cells to estimate the effects of lncRNA-H19 on glioma growth in vivo. RESULTS: Knocking down lncRNA-H19 gene could effectively suppress the proliferation of U251 and U87-MG cells. The knockdown of lncRNA-H19 remarkably inhibited the migration and blocked cycle progressions of U251 and U87-MG cells, yet, no obvious changes were observed in cell apoptosis. Besides, inhibiting lncRNA-H19 expression could attenuate sphere-forming function of U251 and U87-MG cells. Additionally, tumor volume and weight were significantly reduced in rats injected with U251 LV-si-H19 cell line compared to untransfected and negative controls, when survival time was obviously prolonged in U251 LV-si-H19 injection groups. CONCLUSION: LncRNA-H19 gene plays a carcinogenic role in glioma progression via enhancing aggressive behavior of glioma cells.

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Available abstract

BACKGROUND: Glioma is an aggressive adult primary cancer, and is characterized by low cure rate, poor prognosis, and high recurrence. The present study aimed to investigate the effect of lncRNA-H19 gene silencing on glioma cell function. METHODS: lncRNA-H19 interference vector (LV3-si-H19) and negative control vector (LV3-NC) were stably transfected into U251 and U87-MG cells, respectively. Quantitative real-time PCR (qRT-PCR) was performed to investigate the expression of lncRNA-H19. Cell proliferation capacity was tested by adopting cell counting kit (CCK8), and propidium iodide (PI) was used for cell cycle analysis. Meanwhile, flow cytometry (FCM) method was used to investigate cell apoptosis, cell migration capacity was detected via wound healing and transwell experiments, and sphere-forming ability was examined in serum-free suspension culture. Additionally, glioma animal models were conducted through injecting U251 cells to estimate the effects of lncRNA-H19 on glioma growth in vivo. RESULTS: Knocking down lncRNA-H19 gene could effectively suppress the proliferation of U251 and U87-MG cells. The knockdown of lncRNA-H19 remarkably inhibited the migration and blocked cycle progressions of U251 and U87-MG cells, yet, no obvious changes were observed in cell apoptosis. Besides, inhibiting lncRNA-H19 expression could attenuate sphere-forming function of U251 and U87-MG cells. Additionally, tumor volume and weight were significantly reduced in rats injected with U251 LV-si-H19 cell line compared to untransfected and negative controls, when survival time was obviously prolonged in U251 LV-si-H19 injection groups. CONCLUSION: LncRNA-H19 gene plays a carcinogenic role in glioma progression via enhancing aggressive behavior of glioma cells.

Key concepts: Glioma, Cell cycle, Flow cytometry, Gene knockdown, Apoptosis, Cell growth, Gene silencing, Cancer research

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