Expression of Apoptosis and Autophagy Genes in HeLa and Hek 293 Cells under Conditions of Nutrient Deprivation

• Published in: Journal of evolutionary biochemistry and physiology Vol. 59; no. 6; pp. 2304 - 2314
• Main Authors: Trubnikova, A. D., Prokopenko, E. S., Sokolova, T. V., Nadei, O. V., Agalakova, N. I.
• Format: Journal Article
• Language: English
• Published: Moscow Pleiades Publishing 01-11-2023; Springer Nature B.V
• Subjects: Animal Physiology; Apoptosis; Autophagy; BAX protein; Bcl-2 protein; Biochemistry; Biomedical and Life Sciences; Cell culture; Cell viability; Cervical carcinoma; Evolutionary Biology; Experimental Papers; Genes; Life Sciences; Tumor cell lines; Tumor cells; Tumors; HeLa; apoptosis genes; autophagy genes; HEK 293; nutrient deprivation
• ISSN: 0022-0930; 1608-3202
• DOI: 10.1134/S0022093023060315

The goal of the study was a comparing the degree of development of autophagy in the human cervical carcinoma cells of HeLa-V and HeLa-R sublines and non-tumor human embryonic kidney cells HEK 293 under two types of starvation conditions—24- and 48-h culture in serum-free DMEM medium and 4-h incubation in Earle’s minimal medium. The work assessed cell viability using MTT method and the expression of apoptosis (
BCL2
,
BAX
,
CASP3
) and autophagy (
ULK1
,
BECN1
,
ATG5
,
ATG14
,
MAP1LC3B
) genes using real-time PCR. Cultivation under serum starvation and in Earl’s medium resulted in a significant decrease in the viability of HEK 293 cells, but had no influence on HeLa-V and HeLa-R cells. In the tumor cells of both lines, the expression of anti-apoptotic gene
BCL2
increased, while in HEK 293 cells the
BCL2
/
BAX
ratio decreased and
CASP3
gene was activated. In HeLa-V and HeLa-R cells, nutrient deprivation stimulated various combinations of genes
ULK1
,
BECN1
,
ATG5
and
ATG14
implicated in the initial stages of autophagy, but none of the treatments affected the expression of
MAP1LC3B
gene. In HEK 293 cells, serum starvation led to increase in expression level of
BECN1
,
ATG5
,
ATG14
and
MAP1LC3B
genes. Thus, stimulation of autophagy in HeLa cells, especially HeLa-R, prevents the development of apoptosis, while in HEK 293 cells the processes of apoptosis and autophagy occur in parallel. Culture in the serum-free DMEM for 48 h appears to be most effective way to induce autophagy in tumor cell lines and, accordingly, the most suitable model for studying the role of autophagy in the development of their resistance to apoptotic pathway of death.