Inhibition of Human Cervical Cancer Hela Cell Line by Meat-Derived Lactic Acid Bacteria of Lactobacillus plantarum IIA-1A5 and Lactobacillus acidophilus IIA-2B4.

<b>Background and Objective:</b> Two Indonesian lactic acid bacteria of<i> L. plantarum </i>I IA-1A5 and <i>L. acidophilus </i>IIA-2B4 were previously isolated from beef with some functional probiotic properties. Nevertheless, the possibility of these strains to have anticancer activity remains unknown. Current study aimed to evaluate the inhibitory properties of intra-and extracellular protein extracts of these two strains against cervical cancer HeLa cells. <b>Materials and Methods:</b> The intracellular and extracellular proteins extract from <i>L. plantarum </i>IIA-1A5 and <i>L. acidophilus </i>IIA-2B4 were collected and designated as IP-LP, IP-LA, EP-LP and EP-LA, respectively. The effect of these extracts on the viability and morphology of HeLa cells were observed using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay and confocal microscopy, respectively. <b>Results:</b> Both IP-LP and IP-LA inhibited HeLa cells in a concentration-dependent manner, with IC₅₀ values of 352.62 and 120.97 µg mL¹, respectively. Meanwhile, the inhibition activity was also observed for EP-LP and EP-LA, <i>albeit</i> very low. The inhibition effect was also confirmed by morphological analysis under confocal electron microscopy which showed the changes in the cell shapes and numbers. <b>Conclusion:</b> Altogether, for the first time this study proposed that the probiotic isolated from Indonesian beef are promising to inhibit cancer cell lines.

Feasibility of Qualitative Testing of BCR-ABL and JAK2 V617F in Suspected Myeloproliperative Neoplasm (MPN) Using RT-PCR Reversed Dot Blot Hybridization (RT-PCR RDB).

BACKGROUND: Defining the presence of BCR-ABL transcript in suspected myeloproliferative neoplasm is essential in establishing chronic myeloid leukemia. In the absence of BCR-ABL, the conventional diagnostic algorithm recommends JAK2 V617F mutation testing to support diagnosis of other MPN diseases such as polycythemia vera, essential thrombocythemia, and primary myelofibrosis. In certain cases of thrombocythemia, simultaneous upfront testing of both BCR-ABL and JAK2 may be desirable. We wanted to test the feasibility of multiplex detection of BCR-ABL transcript variants and JAK2 V617F mutation simultaneously using the reverse transcriptase polymerase chain reaction (RT-PCR)-based reverse dot-blot hybridization (RDB) method. MATERIAL AND METHODS: Separate biotinylated RT-PCR primers were designed to amplify specific BCR-ABL transcripts and JAK2 V617F mutant alleles. Specific hybridization of RT-PCR products with arrays of membrane-bound probes followed by colorimetric development would allow simultaneous visualization of BCR-ABL and/or JAK2 mutant transcripts in a given specimen. To validate the RDB method, we used cDNA specimens previously referred to our laboratory for routine clinical testing of BCR-ABL and/or JAK2. RESULTS: The limit of detection or analytical sensitivity of the RDB method using cDNA specimens was 0.5% and 6.25% in detecting BCR-ABL and JAK2 mutant transcripts, respectively. The diagnostic specificity and sensitivity to detect BCR-ABL and JAK2 were 100% and 92.3% (N = 38); and 100% and 100% (N = 27), respectively. RDB also detected BCR-ABL transcripts in 22% of JAK2 V617F mutation-positive samples (N = 14). CONCLUSIONS: RT-PCR RDB is a promising qualitative multiplex method to detect BCR-ABL and JAK2 mutant transcripts simultaneously.