PMC

POL cells enhance the migration and invasion abilities of OL cells by delivering exosomal miRNA‐487a‐5p. (A, B) Expression differences of miRNA‐487a‐5p in OL and POL cells (A) and in their respective exosomes (B) detected by RT‐qPCR; n = 3. (C, D) miRNA‐487a‐5p expression in OL cells that were treated with POL exosomes (C), or treated with POL and POL‐miRNA‐487a‐OE exosomes (D). (E) Imaging changes of POL‐exo uptake by OL cells. POL cell‐derived exosomes were labeled with PKH26 (red); the nuclei of OL cells were labeled with DAPI (blue) (bar = 100 μm); n = 3. (F, G) Expression validation of miRNA‐487a‐5p in POL‐Vetor and POL‐miRNA‐487a‐OE cells (F) and in their respective exosomes (G); n = 3. (H, I) The POL‐miRNA‐487a‐OE exosomes promoted the (H) migration and (I) invasion abilities of OL cells (bar = 60 μm); n = 3. (J, K) miRNA‐487a‐5p‐inhibitor treatment reversed the increase in (J) migration and (K) invasion of OL cells that treated with POL‐miRNA‐487a‐OE exosomes (bar = 60 μm); n = 3. (L, M) GW6849 (20 μm) treatment reversed the increase in (L) migration and (M) invasion of OL cells that co‐cultured with POL cells (bar = 60 μm); n = 3. (N) Confirmation of the efficiency of miRNA‐487a‐5p‐mimic treatment on miRNA‐487a‐5p expression; n = 3. (O) Expression of miRNA‐487a‐5p in OL‐Vetor and OL‐miRNA‐487a‐OE cells; n = 3. (P, Q) miRNA‐487a‐5p‐mimic treatment enhanced the (P) migration and (Q) invasion abilities of OL cells (bar = 60 μm); n = 3. NC, negative control; OE, overexpression; OL, oligometastatic cell line; POL, polymetastatic cell line. n = 3: three times a particular experiment was replicated. Error bars indicated SD. *P < 0.05, **P < 0.01, ***P < 0.001 by t‐test.

Effect of OL on apoptosis of Tu686 and CAL-27 cells. A Flow cytometry apoptosis diagrams for different treatment groups with Tu686 (left lane) and CAL-27 (right lane) cells. The dose of 25 µg/mL of OL was selected for flow cytometry. The cells treated with PBS, 25 µg/mL OL, 10 ng/mL TGF-β1, and 10 ng/mL TGF-β1 + 25 µg/mL OL, were named control, OL, TGF-β1, and TGF-β1 + OL groups, respectively. B Comparison column chart of the apoptosis ratio of the four groups with Tu686 cells. Total apoptosis ratio (including early and late) was detected for each group. *p <0.05 vs. control or TGF-β1 group (tests were repeated for 3 times)

Pancherz analysis. OL: line drawn tangent to disto-buccal cusp of maxillary first permanent molar and bisecting the incisal vertical overbite. OLp: a line perpendicular to OL through sella. Sp: (subspinale) deepest point on the anterior contour of the maxillary alveolar projection by tangent perpendicular to OL. Ii (incision inferius): the incisal tip of the most prominent mandibular central incisor. Is (incision superius): the incisal tip of the most prominent maxillary central incisor. Ms (molar superius): the mesial contact point of the maxillary permanent first molar determined by a tangent to OL. Mi (molar inferius): the mesial contact point of the mandibular permanent first molar determined by a tangent perpendicular to OL. Pg (pogonion): the most prominent point on the bony chin determined by a tangent perpendicular to OL. (1) Sp/OLp. (2) Li/OLp. (3) Ms/OLp. (4) Mi/OLp. (5) Pg/OLp.

a, OL Ca²⁺ response in control condition and with Cd²⁺ (50 µM) revealed a slight increase in the 50 Hz-evoked Ca²⁺ rise (n = 54 cells from 4 mice, p = 0.0913 two-sided paired t-test). b, OL Ca²⁺ response with Ni²⁺ (200 µM) revealed a slight decrease in the 50 Hz-evoked Ca²⁺ surge by 11 ± 5% (n = 60 cells from 4 mice, p = 0.0320 two-sided paired t-test). c, OL Ca²⁺ response with Nifedipine (50 µM) revealed a slight decrease in the 50 Hz-evoked Ca²⁺ surge by 18 ± 9% (n = 39 cells from 3 mice, p = 0.0423 two-sided paired t-test). d, OL Ca²⁺ response with Benidipine (10 µM) revealed no overt change in the 50 Hz-evoked Ca²⁺ surge (n = 56 cells from 3 mice, p = 0.2741 two-sided paired t-test). e, OL Ca²⁺ response with RuR (10 µM) revealed no significant changes in the 50 Hz-evoked Ca²⁺ surge (n = 71 cells from 4 mice, p = 0.1405 two-sided paired t-test). Traces represent mean ± SEM. Boxplots show median (line), quartiles (box bounds), mean (+), and 5-95 percentiles (whiskers).