Abstract:

Objective To explore the effects of knocking down glycine cleavage system H protein（GCSH） on proliferation， apoptosis， oxidative stress and migration of gastric cancer SNU-1 cells in vitro. Methods SNU-1 cells were cultured in vitro and divided into control group （no transfection）， negative control group （transfection of negative control siRNA） and GCSH knockdown group （transfection of GCSH siRNA）. Quantitative PCR was used to detect the knockdown effect. Immunofluorescence was used to observe the morphology of cells in each group. CCK-8 was used to test the proliferation of SNU-1 cells. Flow cytometry was used to detect the apoptosis and oxidative stress level， and scratch test was used to detect the cell migration. Results Quantitative PCR experiment showed that the relative expression levels of GCSH in the control group， negative control group and GCSH knockdown group were 1.29±0.16， 1.36±0.17 and 0.32±0.04， respectively （F=90.32， P<0.001）. There was no significant difference between the control group and negative control group （P=0.497）. Compared to the negative control group， the GCSH knockdown group was significantly decreased （P<0.001）. Immunofluorescence experiment showed no significant difference in the morphology of cells among the groups. The CCK-8 experiment results showed that the cell proliferation activities of the control group， negative control group and GCSH knockdown group were 2.63±0.12， 2.61±0.14， 2.45±0.14， respectively （F=6.35， P=0.005）. There was no significant difference between the control group and negative control group （P=0.751）， and the GCSH knockdown group significantly decreased compared to the negative control group （P=0.011）. The results of flow cytometry showed that the early stage apoptosis rates of SNU-1 cells in the control group， negative control group and GCSH knockdown group were （13.38±0.45）%， （12.86±0.65）%， （20.04±3.61）%， respectively （F=15.37， P<0.001）. There was no significant difference between the control group and negative control group （P=0.559）. Compared to the negative control group， the GCSH knockdown group significantly increased （P=0.002）. The late stage apoptosis rates of the three groups were （2.21±0.25）%， （2.68±0.45）%， （5.67±1.67）%， respectively （F=18.24， P<0.001）. There was no significant difference between the control group and negative control group （P=0.356）. Compared to the negative control group， the GCSH knockdown group showed a significant increase （P=0.024）. The reactive oxygen species positive rates in the control group， negative control group and GCSH knockdown group were （26.98±8.79）%， （28.27±5.63）%， （48.41±0.94）%， respectively （F=22.56， P<0.001）. There was no significant difference between the control group and negative control group （P=0.950）. Compared to the negative control group， the GCSH knockdown group significantly increased （P<0.001）. The cell migration rates of the control group， negative control group and GCSH knockdown group were （48.29±5.79）%， （51.66±2.29）%， （14.01±1.56）%， respectively （F=148.80， P<0.001）. There was no significant difference between the control group and negative control group （P=0.328）. Compared with the negative control group， the GCSH knockdown group significantly decreased （P<0.001）. Conclusion Knock down of GCSH gene can inhibit the proliferation and migration， increase cell apoptosis rate and oxidative stress of SNU-1 cells in vitro. GCSH gene may be a potential target for the treatment of gastric cancer.

Key words: Gastric neoplasms, GCSH, Apoptosis, Cell migration