https://orcid.org/0000-0003-0283-0801
Figures & data
Table 1. Primers used for qRT-PCR in this study.
Figure 1. The expressions of circ_0003747 and PLCD3 are increased, miR-338-3p expression is reduced in TC cells.
(a-c) The levels of circ_0003747, miR-338-3p, and PLCD3 were measured using qRT-PCR. (d, e) PLCD3 protein expression was assessed by western blot. (f) The molecular characteristic structure of circ_0003747 was determined by Sanger sequencing. (g) The stability of circ_0003747 was detected by qRT-PCR after the RNase R treatment. *P < 0.05, **P < 0.01, ***P < 0.001.
Figure 2. Circ_0003747 knockdown suppresses TC cells progression.
(a, b) The levels of circ_0003747 and PLCD3 after transfection with sh-circ_0003747 were assessed using qRT-PCR. (c) PLCD3 protein level in TC cells with circ_0003747 knockdown was detected by western blot. (d, e) Cell viability and proliferation were evaluated by MTT and colony formation assays. (f) The wound healing assay was conducted to detect cell migration activity. (g) Cell invasion activity was measured by transwell assay. (h) Cell apoptosis after transfection was monitored by flow cytometry. *P < 0.05, **P < 0.01, ***P < 0.001.
Figure 3. Circ_0003747 negatively regulates miR-338-3p expression in TC cells.
(a) The predicted correlation between circ_0003747 and miR-338-3p was analysed using the CircInteractome database. (b) The binding relationship between circ_0003747 and miR-338-3p was verified by a dual-luciferase reporter assay. (c) The expression of miR-338-3p in TC cells was detected by qRT-PCR. (d, e) Cell viability and proliferation were assessed. (f) Cell invasion activity was measured by transwell assay. (g) Cell apoptosis of TC cells after transfection was monitored using flow cytometry. *P < 0.05, **P < 0.01, ***P < 0.001.
Figure 4. MiR-338-3p enrichment inhibits TC cell progression through targeting PLCD3.
(a) The prediction of the binding site between miR-338-3p and PLCD3 was illustrated using StarBase. (b) The binding relationship between miR-338-3p and PLCD3 was verified by a dual-luciferase reporter assay. (c) The expressions of miR-338-3p and PLCD3 after miR-338-3p mimic transfection were detected using qRT-PCR. (d, e) PLCD3 mRNA and protein expressions were assessed using qRT-PCR and western blot. (f, g) Cell viability and proliferation ability were measured by MTT and colony formation assay. (h) Cell invasion activity was assessed by transwell assay. (i) Cell apoptosis of each group was monitored by flow cytometry. *P < 0.05, **P < 0.01, ***P < 0.001.
Figure 5. Overexpression of PLCD3 reverses the effects of circ_0003747 knockdown.
(a) The mRNA level of PLCD3 with sh-circ_0003747 or PLCD3 transfection was measured by qRT-PCR. (b) PLCD3 protein expression was detected using western blot. (c, d) Cell viability and proliferation ability were measured by MTT and colony formation assay. (e) Cell invasion activity was assessed by transwell assay. (f) Cell apoptosis was monitored by flow cytometry. **P < 0.01, ***P < 0.001.
Figure 6. Circ_0003747 knockdown inhibits tumour growth in nude mice.
(a, b) The tumour growth was assessed by detecting the tumour size and weight. (c) The expressions of miR-338-3p and PLCD3 were detected using qRT-PCR assay. (d) The protein level of PLCD3 in vivo was measured using western blot. (e) The PLCD3 level was determined by immunohistochemistry assay. **P < 0.01, ***P < 0.001.
Supplemental material