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[Circ_0000263 improves radiosensitivity of Hela cells by inhibiting the activity of telomerase protein through miR-338-3p/TERT].
Wang, C; Huo, Y K; Li, M Y; Li, C; Shen, X H; Wang, S J; Liu, Y F; Jiang, Z X.
Afiliación
  • Wang C; Department of Hematology, the First Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, China.
  • Huo YK; Department of Hematology, the First Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, China.
  • Li MY; Department of Hematology, the First Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, China.
  • Li C; Department of Hematology, the First Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, China.
  • Shen XH; Department of Hematology, the First Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, China.
  • Wang SJ; Department of Hematology, the First Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, China.
  • Liu YF; Department of Hematology, the First Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, China.
  • Jiang ZX; Department of Hematology, the First Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, China.
Zhonghua Zhong Liu Za Zhi ; 46(7): 676-685, 2024 Jul 23.
Article en Zh | MEDLINE | ID: mdl-39034803
ABSTRACT

Objective:

To explore the effect and molecular mechanism of circ_0000263 on HeLa cell activity, apoptosis, telomerase activity, and radiosensitivity.

Methods:

The Hela cells were divided into si-NC, si-circ, vector, circ_0000263, anti-NC, anti-miR-338-3p, miR-NC, miR-338-3p, si-circ+anti-NC, si-circ+ anti-miR-338-3p, si-circ+vector, si-circ+TERT, sh-NC, sh-circ groups. Reverse transcription-quantitative real-time polymerase chain reaction (RT-qPCR) was used to detect the expressions of circ_0000263 and miR-338-3p. Cell clone formation array was used to detect cell survival; cell counting kit-8 (CCK-8) to detect cell proliferation; flow cytometry to detect apoptosis; western blot method to detect the expressions of proliferating cell nuclear antigen (PCNA), Cleaved-casp3, telomerase reverse transcriptase (TERT) proteins; double luciferase assay to detect the targeting relationships of circ_0000263 and miR-338-3p, miR-338-3p and TERT; telomere repeat amplification enzyme linked immunosorbent assay (TRAR-ELISA) to detect telomerase activity.

Results:

Circ_0000263 was highly expressed in Hela cells, miR-338-3p was low expressed, and TERT was highly expressed; circ_0000263 was also highly expressed in Hela cells treated with radiation (P<0.05). Knockdown of circ_0000263 inhibited the clone formation and cell proliferation ability of HeLa cells, and enhanced the radiosensitivity and apoptosis of HeLa cells. In contrast, knockdown of circ_0000263 decreased PCNA protein expression level and enhanced Cleaved-casp3 protein expression level in HeLa cells (P<0.05). The apoptosis rate in the si-circ group was (13.19±1.12)%, which was higher than (6.80±0.62)% of si-NC group (P<0.05). The apoptosis rate in the si-circ+4 Gy group was (24.82±1.57)%, which was higher than (17.00±0.96)% of si-NC+4 Gy group (P<0.05). Circ_0000263 targeted regulated miR-338-3p, and miR-338-3p targeted regulated TERT. MiR-338-3p was lowly expressed in HeLa cells, and knockdown of circ_0000263 elevated miR-338-3p expression level in HeLa cells. Circ_0000263 regulated TERT expression and inhibited telomerase activity through miR-338-3p. MiR-338-3p/TERT can restore the effect of circ_0000263 on the radiosensitivity of Hela cells. The apoptosis rate in the si-circ+anti-NC group was (27.37±0.89)%, which was higher than (18.22±1.18)% of the si-circ+anti-miR-338-3p group (P<0.05). The apoptosis rate in the si-circ+vector group was (27.55±0.48)%, which was higher than (20.10±0.68)% of si-circ+TERT group (P<0.05). After 72 hours of radiation by 4 Gy, the cell survival fraction of si-circ+anti-NC group was 0.41±0.02, which was lower than 0.66±0.03 of the si-circ+anti-miR-338-3p group (P<0.05); the cell survival fraction of si-circ+vector group was 0.42±0.05, which was lower than 0.70±0.03 of si-circ+TERT group (P<0.05).

Conclusion:

Inhibiting the expression of circ_0000263 supresses the proliferation of Hela cells by regulating miR-338-3p/TERT, promotes apoptosis, inhibits telomerase activity, increases the radiosensitivity of cancer cells, and provides a theoretical basis for improving the radiosensitivity of Hela cells.
Asunto(s)

Texto completo: 1 Bases de datos: MEDLINE Asunto principal: Tolerancia a Radiación / Apoptosis / Telomerasa / MicroARNs / Proliferación Celular Límite: Humans Idioma: Zh Revista: Zhonghua Zhong Liu Za Zhi Año: 2024 Tipo del documento: Article País de afiliación: China

Texto completo: 1 Bases de datos: MEDLINE Asunto principal: Tolerancia a Radiación / Apoptosis / Telomerasa / MicroARNs / Proliferación Celular Límite: Humans Idioma: Zh Revista: Zhonghua Zhong Liu Za Zhi Año: 2024 Tipo del documento: Article País de afiliación: China