The Relativity Between Staphylococcus Aureus L-forms And Carcinoma Of Gallbladder

Objectives: To investigate the relationship between the S.aureus L-forms and Carcinoma of Gallbladder.Methods:1. Using penicillin slip to induce the stable L-forms of S.aureus. 2. Under the action of the S.aureus L-forms, with the ratio of S.aureus L-forms and GBC-SD was 100:1, we want to explore the conglutination of the S.aureus L-forms to the GBC-SD by the light microscope, and whether the S.aureus L-forms can enter the GBC-SD by PCR reaction and electron microscope. 3. Under the action of different proportion between S.aureus L-forms and GBC-SD such as 1:10,1:1,10:1,100:1 for 1,2 or 4 days, we want to explore the effect on the proliferation of GBC-SD of the stable L-forms of S.aureus by the MTT and Proliferating Cell Nuclear Antigen(PCNA).In addition, under the condition of 100:1, we introduce the hours of 3,6,12,24,48 hours separately to explore the effect on the apoptosis of GBC-SD of the stable L-forms of S.aureus by the Flow Cytometry(FCM) and DNA Ladder.4. Using the Human U133 Plus 2.0 gene chip to detect gene contrast after the action of the stable L-forms of S.aureus, and using the RT-PCR to prove the gene contrast within the gene chip.ResuLts: 1. The stable L-forms of S.aureus were induced successfully after 18 generation and did not reply after 3 generation without penicillin. 2. We observed that the S.aureus L-forms stick on the GBC-SD and entered the GBC-SD. 3. The stable L-forms of S.aureus promoted the proliferation of GBC-SD by the MTT and PCNA when the GBC-SD was effected by the stable L-forms of S.aureus on 4 days(P<0.05),and the growth rates were 16.7%,19.2%,13.7%,24.1% with the ratio of 1:10,1:1,10:1,100:1,the growth rates were not different between the different ratio of S.aureus L-forms and GBC-SD(P>0.05). Forthermore, the stable L-forms of S.aureus did not cause the apoptosis of GBC-SD by the Flow Cytometry(FCM) and DNA Ladder when the GBC-SD was effected by the stable L-forms of S.aureus for 3,6,12,24,48 hours, the collator of the S.aureus and the instable L-forms caused the apoptosis of GBC-SD, and the apoptosis caused by the instable L-forms of S.aureus was lower than the S.aureus. 4. Twenty gene were changed when the GBC-SD was effect by the stable L-forms of S.aureus by the gene chip, the up-regulation gene was four and the Genebank ID was AK024454,AK021539,AI871627,AW299815. The down-regulation gene was sixteen and the Genebank ID was AI985407,BC041998,BC035193,NM₀01393,L31792,M81778,AF006623,NM₀25134,NM₀23921,AW117584,AK023065,AK025063,R44298,BE463757,AW873604,AA018404. We proved that the up-regulation gene of EGF-Like moduLe-containing mucin-Like hormone receptor-Like 2 (EMR2) and the up-regulation gene of extraceLLuLar matrix protein 2(ECM2) was match with the gene chip by RT-PCR(P<0.05).ConcLusions:The S.aureus L-forms can enter the GBC-SD, the stable L-forms of S.aureus can promote the proliferation of GBC-SD, the S.aureus and its instable L-forms can cause the apoptosis of GBC-SD, the gene chip indicate that the stable L-forms of S.aureus can cause the up-regulation of EMR2 and the down-reguLation of ECM2 of GBC-SD, which indicate that there is association between the S.aureus L-forms and the carcinogenesis,carcinomatous development of Gallbladder.