Effect of lncRNA HULC knockdown on rat secreting pituitary adenoma GH3 cells

Pituitary adenoma is one of the most common tumors in the neuroendocrine system. This study investigated the effects of long non-coding RNAs (lncRNAs) highly up-regulated in liver cancer (HULC) on rat secreting pituitary adenoma GH3 cell viability, migration, invasion, apoptosis, and hormone secretion, as well as the underlying potential mechanisms. Cell transfection and qRT-PCR were used to change and measure the expression levels of HULC, miR-130b, and FOXM1. Cell viability, migration, invasion, and apoptosis were assessed using trypan blue staining assay, MTT assay, two-chamber transwell assay, Guava Nexin assay, and western blotting. The concentrations of prolactin (PRL) and growth hormone (GH) in culture supernatant of GH3 cells were assessed using ELISA. The targeting relationship between miR-130b and FOXM1 was verified using dual luciferase activity. Finally, the expression levels of key factors involved in PI3K/AKT/mTOR and JAK1/STAT3 pathways were evaluated using western blotting. We found that HULC was highly expressed in GH3 cells. Overexpression of HULC promoted GH3 cell viability, migration, invasion, PRL and GH secretion, as well as activated PI3K/AKT/mTOR and JAK1/STAT3 pathways. Knockdown of HULC had opposite effects and induced cell apoptosis. HULC negatively regulated the expression of miR-130b, and miR-130b participated in the effects of HULC on GH3 cells. FOXM1 was a target gene of miR-130b, which was involved in the regulation of GH3 cell viability, migration, invasion, and apoptosis, as well as PI3K/AKT/mTOR and JAK1/STAT3 pathways. In conclusion, HULC tumor-promoting roles in secreting pituitary adenoma might be via down-regulating miR-130b, up-regulating FOXM1, and activating PI3K/AKT/mTOR and JAK1/STAT3 pathways.

Effects of LncRNA HULC on radiosensitivity of osteosarcoma cells / 中华放射医学与防护杂志

Objective To investigate the effect of LncRNA HULC on radiosensitivity of osteosarcoma cells. Methods Osteosarcoma cells OS732 was infected by shRNA HULC lentivirus, and the interference effect was determined by qRT-PCR. Osteosarcoma cells infected with shRNA HULC lentivirus were irradiated with 8 Gy X-rays. MTT, PI monochrome staining and Annexin V-FITC/PI double staining were used to detect cell proliferation, cell cycle and apoptosis, respectively. Western blot was used to detect the protein levels of p21, Cyclin D1, C-Caspase-3 and Cyt-C in cytoplasm and mitochondria. Plate cloning assay was used to evaluate cell radiosensitivity. Results The expression of HULC in osteosarcoma cells was significantly down-regulated by shRNA HULC lentivirus infection. Down-regulation of HULC or irradiation inhibited osteosarcoma cell proliferation [(100. 00±9. 65)％ vs. (71. 36±5. 27)％, (63. 48± 5. 93)％, t=4. 512, 5. 585, P<0. 05 ] , blocked cell cycle [ ( 50. 15 ± 5. 14 )％ vs. ( 62. 35 ± 4. 22 )％, (66. 05±5. 23)％,t=3. 177,3. 756,P<0. 05], induced cell apoptosis [(2. 98±0. 23)％ vs. (22. 61± 3. 26)％, (26. 14±2. 81)％,t=8. 898,10. 498,P<0. 05], promoted the expressions of p21 and Cyclin D1 in cells, down-regulated the level of C-Caspase-3 protein, increased the level of Cyt-C protein in cytoplasm, and down-regulated the level of Cyt-C protein in mitochondria. Downregulation of HULC combined with irradiation yield much more effects on cell proliferation inhibition [ ( 71. 36 ± 5. 27 )％, (63.48±5.93)％ vs. (49.32±5.76)％, t=4.890, 2.967, P<0.05], cell cycle arrest [(62.35± 4. 22)％, (66. 05±5. 23)％ vs. (77. 17±7. 54)％, t=2. 983, 2. 106, P<0. 05], apoptosis induction [(22. 61±3. 26)％, (26. 14±2. 81)％ vs. (36. 21±3. 26) ％,t=6. 164, 4. 564, P<0. 05] and the expressions of p21, Cyclin D1, C-Caspase-3 and Cyt-C in osteosarcoma cells. The radiosensitization ratio of down-regulation of HULC was 1. 432. Conclusions Down-regulation of HULC enhances radiosensitivity of osteosarcoma cells, which may be related to cell cycle arrest and apoptosis induction.