Lnc-GULP1–2:1 affects granulosa cell proliferation by regulating COL3A1 expression and localization

Abstract

Backgrounds Long non-coding RNA is a novel group of non-protein coding transcripts over 200 nt in length. Recent studies have found that they are widely involved in many pathological and physiological processes. In our previous study, we found that lnc-GULP1–2:1 was significantly down-regulated in the ovarian cortical tissue of patients with primary ovarian insufficiency and predicted that lnc-GULP1–2:1 has a regulatory effect on COL3A1. Results In this study, we found that lnc-GULP1–2:1 was mainly localized in the cytoplasm of luteinized granulosa cells. The expression of lnc-GULP1–2:1 was lower in patients with diminished ovarian reserve but substantially elevated in patients with polycystic ovary syndrome. Overexpression of lnc-GULP1–2:1 in KGN cells significantly inhibited cell proliferation, likely through cell cycle related genes CCND2 and p16. Moreover, lnc-GULP1–2:1 expression was positively correlated with the level of COL3A in luteinized granulosa cells from patients with different ovarian functions as well as in multiple cell lines. Overexpression of lnc-GULP1–2:1 in KGN cells promoted the expression of COL3A1 and its translocation into the nucleus. Consistently, silencing COL3A1 in KGN cells also significantly inhibited cell proliferation. Conclusions Lnc-GULP1–2:1 affects the proliferation of granulosa cells by regulating the expression and localization of COL3A1 protein, and may participate in the regulation of ovarian follicle development. This study will provide new insight into molecular mechanisms underlying ovarian follicular development, which will help generate novel diagnostic and therapeutic strategies for diseases related to ovarian follicular development disorders.

Long non-coding RNA Loc105611671 promotes the proliferation of ovarian granulosa cells and steroid hormone production upregulation of CDC42

“…For example, lncRNA ZNF674-AS1 affects GC proliferation by interacting with ALDOA ( 31 ). lnc-GULP1-2:1 was also found to bind directly to COL3A1 and affect GC proliferation by regulating COL3A1 expression and localization ( 32 ). In this study, we used RNA pull-down, mass spectrometry, and dual-luciferase gene assay experiments to further reveal that CDC42 directly binds to and interacts with Loc105611671 in GCs.…”

Expression of genes regulating cell division in porcine follicular granulosa cells

“…It is worth noting the role of COL3A1, an ECM component protein in folliculogenesis, whose expression has been demonstrated within the ovary in both cattle and pigs, mono- and polyovulatory animals [ 73 ]. COL3A1 expression in PCOS [ 74 ] as well as POI [ 75 ] suggest that Lnc-GULP1-2:1 (long non-coding RNA) could be used to alter COL3A1 expression, treating it as a therapeutic target. Additionally, COL3A1 has been shown to be a biomarker for ovarian cancers [ 76 ] and also its elevated expression reduces the effects of anticancer drugs in vitro [ 77 ].…”

“…Many glycoproteins found in the ECM of granulosa cells have been described to influence the occurrence of diseases within the ovary, including PCOS and POI [ 107 – 109 ]. Interestingly, several genes encoding cytoskeleton-related proteins have also been described in relation to PCOS, including: TPM2, VCAM-1 [ 47 , 70 ], and in the case of COL3A1 also in the aspect of POI [ 74 , 75 ]. Given some ability to modulate cytoskeleton composition with actin-binding proteins [ 41 , 110 ], it should be possible to influence ECM composition.…”

Effect of Human Amniotic Epithelial Stem Cell Transplantation on Preterm Premature Rupture of Fetal Membrane Using the Amniotic Pore Culture Technique in vitro

“…Isolated hAESCs were fixed with 4% paraformaldehyde for 10 min and washed three times with PBS. The experimental protocols were learned from the previous papers [ 20 , 21 , 22 ]. The cells were then incubated with antibodies against cluster of differentiation 19 (CD19), CD34, CD73, CD90 (Thy1), CD105 (endoglin), CD324 (E-cadherin), stage-specific embryonic antigen 4 (SSEA4), octamer-binding transcription factor 3/4 (Oct3/4), SRY-box transcription factor 2 (Sox2), Nanog (BD Biosciences, San Jose, CA, USA), and human leukocyte antigen (HLA) class I or II (Ancell, MN, USA) for 40 min on ice after a blocking procedure.…”