Zhongguo shi yan xue ye xue za zhi · 2026
[LncRNA PVT1 Regulates the MiR-20a-5p/HMGA2 Signaling Axis to Affect Proliferation and Apoptosis of Diffuse Large B-Cell Lymphoma Cells].
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Xu Xiao-Wei, Adina Wutikuer, Zhang Hong-Li, Shi Yu-Wei
Аннотация
OBJECTIVE: To investigate the effect of long non-coding RNA (lncRNA) PVT1 on proliferation and apoptosis of diffuse large B-cell lymphoma (DLBCL) cells and the underlying mechanisms.
METHODS: qRT-PCR was used to detect the expression of PVT1 in human B lymphoblastic cells IM-9 and DLBCL cells OCI-Ly1, OCI-Ly3, OCI-Ly4, and OCI-Ly10. OCI-Ly1 cells were divided into four groups: si-NC (negative control), si-PVT1, si-PVT1+miR-20a-5p inhibitor, and si-PVT1+miR-20a-5p inhibitor+si-HMGA2. OCI-Ly3 cells were divided into four groups: Control, PVT1, PVT1+miR-20a-5p mimic, and PVT1+miR-20a-5p mimic+HMGA2. Dual-luciferase reporter assay was performed to verify the targeting relationship between PVT1 and miR-20a-5p, as well as the targeting relationship between miR-20a-5p and HMGA2. CCK-8 assay was used to detect cell proliferative capacity. Flow cytometry was used to detect cell apoptosis, and Western blot was used to detect the expression of HMGA2 protein.
RESULTS: The expression of PVT1 in OCI-Ly1, OCI-Ly3, OCI-Ly4, and OCI-Ly10 cells was significantly higher than that in IM-9 cells. Dual-luciferase reporter assay confirmed that miR-20a-5p could bind specifically to PVT1, and HMGA2 is a target of miR-20a-5p. In OCI-Ly1 cells, the si-PVT1 group exhibited significantly reduced cell proliferation and HMGA2 protein expression (P <0.05) and markedly increased apoptosis (P <0.001) compared with the si-NC group; The si-PVT1 + miR-20a-5p inhibitor group exhibited significantly higher proliferation (P <0.01) and substantially lower apoptosis (P <0.001) than the si-PVT1 group; Furthermore, the si-PVT1 + miR-20a-5p inhibitor + si-HMGA2 group displayed significantly decreased proliferation (P <0.01) and dramatically elevated apoptosis (P <0.001) when compared with the si-PVT1 + miR-20a-5p inhibitor group. In OCI-Ly3 cells, the PVT1 group displayed significantly enhanced proliferation and elevated HMGA2 protein expression (P <0.05) with markedly reduced apoptosis (P <0.001) compared to the Control group. Conversely, the PVT1 + miR-20a-5p mimic group showed dramatically impaired proliferation (P <0.001) and substantially increased apoptosis (P < 0.001) versus the PVT1 group. Notably, the PVT1 + miR-20a-5p mimic + HMGA2 group rescued proliferation (P <0.05) and suppressed apoptosis (P <0.001) relative to the PVT1 + miR-20a-5p mimic group.
CONCLUSION: PVT1 promotes the proliferation and inhibits the apoptosis of DLBCL cells by regulating the miR-20a-5p/HMGA2 signaling axis.
METHODS: qRT-PCR was used to detect the expression of PVT1 in human B lymphoblastic cells IM-9 and DLBCL cells OCI-Ly1, OCI-Ly3, OCI-Ly4, and OCI-Ly10. OCI-Ly1 cells were divided into four groups: si-NC (negative control), si-PVT1, si-PVT1+miR-20a-5p inhibitor, and si-PVT1+miR-20a-5p inhibitor+si-HMGA2. OCI-Ly3 cells were divided into four groups: Control, PVT1, PVT1+miR-20a-5p mimic, and PVT1+miR-20a-5p mimic+HMGA2. Dual-luciferase reporter assay was performed to verify the targeting relationship between PVT1 and miR-20a-5p, as well as the targeting relationship between miR-20a-5p and HMGA2. CCK-8 assay was used to detect cell proliferative capacity. Flow cytometry was used to detect cell apoptosis, and Western blot was used to detect the expression of HMGA2 protein.
RESULTS: The expression of PVT1 in OCI-Ly1, OCI-Ly3, OCI-Ly4, and OCI-Ly10 cells was significantly higher than that in IM-9 cells. Dual-luciferase reporter assay confirmed that miR-20a-5p could bind specifically to PVT1, and HMGA2 is a target of miR-20a-5p. In OCI-Ly1 cells, the si-PVT1 group exhibited significantly reduced cell proliferation and HMGA2 protein expression (P <0.05) and markedly increased apoptosis (P <0.001) compared with the si-NC group; The si-PVT1 + miR-20a-5p inhibitor group exhibited significantly higher proliferation (P <0.01) and substantially lower apoptosis (P <0.001) than the si-PVT1 group; Furthermore, the si-PVT1 + miR-20a-5p inhibitor + si-HMGA2 group displayed significantly decreased proliferation (P <0.01) and dramatically elevated apoptosis (P <0.001) when compared with the si-PVT1 + miR-20a-5p inhibitor group. In OCI-Ly3 cells, the PVT1 group displayed significantly enhanced proliferation and elevated HMGA2 protein expression (P <0.05) with markedly reduced apoptosis (P <0.001) compared to the Control group. Conversely, the PVT1 + miR-20a-5p mimic group showed dramatically impaired proliferation (P <0.001) and substantially increased apoptosis (P < 0.001) versus the PVT1 group. Notably, the PVT1 + miR-20a-5p mimic + HMGA2 group rescued proliferation (P <0.05) and suppressed apoptosis (P <0.001) relative to the PVT1 + miR-20a-5p mimic group.
CONCLUSION: PVT1 promotes the proliferation and inhibits the apoptosis of DLBCL cells by regulating the miR-20a-5p/HMGA2 signaling axis.