SSRP1 affects the growth and apoptosis of gastric cancer cells through AKT pathway Translated title: SSRP1 utiče na rast i apoptozu ćelija raka želuca kroz AKT signalni put

Abstract

Background

We aimed to determine the SSRP1's potential influence on the apoptosis and proliferation of gastric cancer (GC) cells and its regulatory mechanism.

Methods

SSRP1 expression in GC cells and tissues was detected via quantitative reverse transcription-polymerase chain reaction (qRT-PCR). The interrelation between clinicopathological characteristics of GC patients and SSRP1 expression was analysed via x2 test, and the correlation between SSRP1 expression and overall survival rate was analysed using Kaplan-Meier survival analysis. After the knockdown of SSRP1 in AGS cells, the SSRP1 expression, colony formation ability, cell viability, cell cycle changes, apoptosis rate, and migration and invasion ability were detected through qRT-PCR, colony formation assay, CCK8 assay, flow cytometry and transwell test, respectively. Finally, the effects of down-regulation of SSRP1 on the expressions of phosphorylated-protein kinase B (p-AKT), B-cell lymphoma-2 (Bcl-2) and Bcl-2 associated X protein (Bax) were explored using Western blotting.

Results

SSRP1 displayed a high expression in GC cells and tissues. SSRP1 expression was closely interrelated to the TNM stage, lymph node metastasis and tumour size. The survival rate of patients was markedly shorter in the high expression group than in the lower expression group. After the knockdown of SSRP1 in cells, the viability and colony formation ability of AGS cells were inhibited. In addition, the cell ratio in the G1 phase was increased, while that in the S phase declined, and the cell invasion and migration were obviously weakened. It was found from Western blotting that the knockdown of SSRP1 could evidently suppress the protein levels of Bcl-2 and p-AKT but promote the protein expression of Bax, indicating that silencing SSRP1 can inhibit the proliferative capacity and increase the number of GC cells through inactivating the AKT signalling pathway.

Conclusions

SSRP1 rose up in GC tissues and cells. Reduction of SSRP1 can inhibit the proliferative capacity and increase the number of GC cells through inactivating the AKT signalling pathway.

Translated abstract

Uvod

Naš cilj je bio da otkrijemo potencijalni uticaj SSRP1 na apoptozu i proliferaciju ćelija raka želuca (GC) i njegov regulatorni mehanizam.

Metode

Ekspresija SSRP1 u GC ćelijama i tkivima je detektovana kvantitativnom lančanom reakcijom polimeraze reverzne transkripcije (qRT-PCR). Odnos između kliničkopatoloških karakteristika pacijenata sa GC i ekspresije SSRP1 je analiziran pomoću Hi-kvadratnog testa (x2), a korelacija između ekspresije SSRP1 i ukupne stope preživljavanja pomoću Kaplan-Meier analize preživljavanja. Nakon obaranja SSRP1 u ćelijama AGS, ekspresija SSRP1, sposobnost formiranja kolonije, vitalnost ćelija, promene ćelijskog ciklusa, stopa apoptoze i sposobnost migracije i invazije su otkriveni pomoću qRT-PCR, testa formiranja kolonije, CCK8 testa, protočne citometrije i transvel testa, redom. Konačno, efekti smanjenja SSRP1 na ekspresije fosforilisane protein kinaze B (p-AKT), B-ćelijskog limfoma2 (Bcl-2) i Bcl-2 pridruženog Ks proteina (Bax) su ispitani pomoću Vestern blota.

Rezultati

SSRP1 je pokazao visoku ekspresiju u GC ćelijama i tkivima. Ekspresija SSRP1 je bila usko povezana sa stadijumom TNM, metastazama u limfnim čvorovima i veličinom tumora. Stopa preživljavanja pacijenata je bila znatno kraća u grupi sa visokom ekspresijom nego u grupi sa nižom ekspresijom. Nakon obaranja vrednosti SSRP1 u ćelijama, došlo je do inhibicije vitalnosti i sposobnosti formi ranja kolonija AGS ćelija. Pored toga, povećana je elijska stopa u G1 fazi, dok je u S fazi opala, dok su invazija i migracija ćelija očigledno oslabljene. Vestern blot metodom je utvđeno da bi obarenje SSRP1 moglo očigledno potisnuti nivo proteina Bcl-2 i p-AKT, ali bi promovisalo ekspresiju proteina Bak, ukazujući da smanjenje SSRP1 može inhibirati proliferativni kapacitet i povećati broj GC ćelija kroz inaktivaciju AKT signalnog puta.

Zaključak

SSRP1 je porastao u tkivima i ćelijama GC. Smanjenje SSRP1 može inhibirati proliferativni kapacitet i povećati broj GC ćelija kroz inaktivaciju AKT signalnog puta.