Correlation And Mechanism Research Between TUBB3,STMN1 And BRCA1 Expression Levels And Drugs Of Gastric Cancer

Objective:To explore the expression of BRCA1, STMN1 and TUBB3 in gastric cancer and para-carcinoma tissue, and analyze their relationship with clinicopathological parameters; and to explore the effect and mechanism of Shenqifuzheng Injection(SFI) and Docetaxel(TXT) on gastric cancer BGC-823 cells.Method:(1) Using immunohistochemistry(Elivision method) to detect the expression of BRCA1, STMN1 and TUBB3 protein in 54 cases of gastric cancer tissues and 20 cases of para-carcinoma tissues.(2) Using CCK8 to detect the effects of different concentrations of SFI(5mg/ml, 10mg/ml, 20mg/ml, 40mg/ml, 80mg/ml) or different concentrations of TXT(3.75mg/L, 7.5mg/L, 15mg/L, 30mg/L, 60mg/L)or TXT(7.5mg/L) combined wit h different concentrations of SFI(5mg/ml, 10mg/ml, 20mg/ml, 40mg/ml, 80mg/ml) for different time(24h,48 h,72h)on the proliferation of BGC-823 cells.(3) Using flow cytometry combined with PI method to detect the effects of different concentrations of SFI(5mg/ml, 20mg/ml, 80mg/ml) or different concent rations of TXT(3.75mg/L, 15mg/L, 60mg/L)or TXT(7.5mg/L) combined with d ifferent concentrations of SFI(5mg/ml, 20mg/ml, 80mg/ml) for 48 h on the cell cycle of BGC-823 cells.(4) Using reverse transcription polymerase chain reaction(RT-PCR) to detect the gene expression of BRCA1, STMN1 and TUBB3 among ten groups, and these groups include BGC-823 cells group(blank control), BGC-823 cells treated with SFI(5mg/ml,20mg/ml 、 80mg/ml) and BGC-823 cells treated with TXT(3.75mg/ml15mg/ml, 30mg/ml) and BGC-823 cells treated with TXT(7.5mg/L)combined with SFI(5mg/ml, 20mg/ml, 80mg/ml) for 48 h.Results:(1)Immunohistochemisrty showed that the positive rate of BRCA1, STMN1 and TUBB3 is higher in gastric carcinoma than para-carcinoma tissue in 54 cases of gastric carcinoma and 20 cases corresponding adjacent tissues; The protein expression of STMN1 is correlated with differentiation(P<0.05). whereas the protein expression of BRCA1 or TUBB3 is not correlated with clinicopathological features(P>0.05). the protein expression of TUBB3 and STMN1 has a certain correlation(P<0.05, r= 0.276).(2) Compared with control group, with the concentration of SFI(5, 10, 20, 40,80mg/ml) or TXT(3.75, 7.5, 15, 30, 60mg/L) increased and the action time(24h, 48 h and 72h) extension, the cell proliferation was significantly inhibited(P<0.05);Compared with control group, with the action time(24h, 48 h and 72h) extension, the cell proliferation was significantly inhibited(P<0.05), but not with the concentration of TXT(7.5 mg/L) combined with SFI(5, 10, 20, 40, 80mg/ml) increased.(3) Different concentration of TXT or SFI could inhibit the proliferation of BGC-823 through blocking G2/M phase or S phase, compared with the control group(P<0.05); Different concentration of TXT combined with SFI could inhibit the proliferation of BGC-823 through blocking G2/M phase, compared with the control group(P<0.05).(4) The gene expression of STMN1 or TUBB3 is lower in experimental groups than control group(P<0.05), while the gene expression of BRCA1 is not significance(P>0.05).Conclusion:(1)STMN1 may be involved in the occurrenceand developmentprocess of gastric cancer; and TUBB3 and BRCA1 is only correlated with occurrenceprocess of gastric cancer.(2)STMN1 could be a molecular marker of gastric cancer, early screening of h igh-risk groups, treatment and follow-up of patients with gastric cancer.