Guizhi Fuling pills inhibit the proliferation, migration and invasion of human cutaneous malignant melanoma cells by regulating the molecular axis of LncRNA TPT1-AS1 / miR-671-5p

Abstract

The current research was aimed to explore the effects of Guizhi Fuling Pills on the proliferation, migration and invasion of human cutaneous malignant melanoma cells and its regulation on the molecular axis of LncRNA TPT1-AS1 / miR-671-5p. Human cutaneous malignant melanoma cells A375 were cultured in vitro and randomly divided into Con group, Guizhi Fuling pills-L group, Guizhi Fuling pills-M group, Guizhi Fuling pills-H group, Guizhi Fuling pills-H + pcDNA group, Guizhi Poria pills-H + pcDNA-TPT1-AS1 group. MTT was used to detect cell proliferation. The Transwell cell test was used to detect cell migration and invasion. qRT-PCR was used to detect the expression of TPT1-AS1 and miR-671-5p. The dual-luciferase report experiment verified the targeting relationship of TPT1-AS1, miR-671-5p. Western blot was used to detect the expression of Ki-67, PCNA, MMP-2 and MMP-9. Compared with Con group, Guizhi Fuling Pills could inhibit cell proliferation, migration and invasion (p<0.05), and also inhibit the expression of Ki-67, PCNA, MMP-2, MMP-9, TPT1-AS1 (p<0.05), promote the expression of miR-671-5p (p<0.05)., and the differences between the indexes of Guizhi Fuling Pill-L group, Guizhi Fuling Pill-M group and Guizhi Fuling Pill-H group were statistically significant (p<0.05). The dual-luciferase report experiment confirmed that TPT1-AS1 could target and bind to miR-671-5p and could regulate the expression and activity of miR-671-5p. Overexpression of TPT1-AS1 could reduce the inhibitory effect of Guizhi Fuling Pills on proliferation, migration and invasion of A375 cells. Guizhi Fuling Pill may reduce the proliferation, migration and invasion of human cutaneous malignant melanoma cells by down-regulating the expression of TPT1-AS1 and up-regulating the expression of miR-671-5p.

MicroRNA‑671‑5p inhibits cell proliferation, migration and invasion in non‑small cell lung cancer by targeting MFAP3L

“…At the molecular level, the present study further demonstrated that 95D cells transfected with miR-671-5p mimics exhibited downregulated expression levels of PCNA, N-cadherin and vimentin, whereas E-cadherin expression levels were upregulated compared with those in the control cells. Guizhi Fuling pills have been reported to inhibit cell proliferation and PCNA expression, which is associated with elevated miR-671-5p expression levels ( 28 ). Tan et al ( 10 ) have demonstrated that the levels of the epithelial marker E-cadherin are upregulated, whereas the levels of vimentin are downregulated in miR-671-5p-transfected MDA-MB-231 cells compared with miR-NC transfected cells.…”

Positive feedback regulation of lncRNA TPT1‐AS1 and ITGB3 promotes cell growth and metastasis in pancreatic cancer

“…Besides, high expression of TPT1‐AS1 was positively correlated with advanced TNM stage and poor differentiation, indicating a poor clinical prognosis of PDAC patients. In addition, previous research showed that TPT1‐AS1 functioned as an oncogene in gastric cancer, colorectal cancer, ovarian cancer, cervical cancer, among others 16–19,39,40 …”

“…Four major binding sites on the promoter region of TPT1-AS1 were predicted In addition, previous research showed that TPT1-AS1 functioned as an oncogene in gastric cancer, colorectal cancer, ovarian cancer, cervical cancer, among others. [16][17][18][19]39,40 Moreover, functional experiments revealed that TPT1-AS1 could promote pancreatic cancer cell proliferation, invasion, migration, and EMT process in vitro and in vivo. These findings were consistent with those of previous studies and suggested that TPT1-AS1 acted as an oncogenic lncRNA in PDAC.…”

Research progress on the role and mechanism of miR-671 in bone metabolism and bone-related diseases

“…LncRNA-miRNA-mRNA regulatory network plays a significant role in cancer's occurrence, development, and prognosis by regulating downstream target genes and proteins (54)(55)(56)(57). Therefore, targeted therapy drugs can inhibit cancer growth by influencing the regulatory network (58,59). In most types of cancer, such as breast cancer, esophageal cancer, nonsmall cell lung cancer, hepatocellular carcinoma, etc., miR-671 acts as a tumor suppressor and is down-regulated in cancer (60)(61)(62)(63)(64)(65)(66)(67)(68)(69).…”