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  1. Silencing E6/E7 Oncoproteins in SiHa Cells Treated with siRNAs and Oroxylum indicum Extracts Induced Apoptosis by Upregulating p53/pRb Pathways
    Sisin NNT, Kong AR, Edinur HA, Jamil NIN, Che Mat NF
    Appl Biochem Biotechnol, 2024 Jul;196(7):4234-4255.
    PMID: 37922032 DOI: 10.1007/s12010-023-04762-w
    E6 and E7 human papillomavirus (HPV) oncoproteins play a significant role in the malignant transformation of infected cervical cancer cells via suppression of tumour suppressor pathways by targeting p53 and pRb, respectively. This study aimed to investigate the anticancer effects of Oroxylum indicum (OI) leaves' methanol extract on SiHa cervical cancer cells. Expression of apoptosis-related proteins (Bcl-2, caspase (cas)-3, and cas-9), viral oncoproteins (E6 and E7), and tumour suppressor proteins (p53 and pRb) were evaluated using western blot analysis before and after E6/E7 small interfering RNAs (siRNAs) transfection. In addition, the E6/E7 mRNA expression levels were assessed with real-time (RT)-PCR. The present study showed that the OI extract effectively hindered the proliferation of SiHa cells and instigated increments of cas-3 and cas-9 expressions but decreased the Bcl-2 expressions. The OI extract inhibited E6/E7 viral oncoproteins, leading to upregulation of p53 and pRb tumour suppressor genes in SiHa cells. Additionally, combinatorial treatment of OI extract and gossypin flavonoid induced restorations of p53 and pRb. Treatment with OI extract in siRNA-transfected cells also further suppressed E6/E7 expression levels and further upregulations of p53 and pRb proteins. In conclusion, OI extract treatment on siRNAs-transfected SiHa cells can additively and effectively block E6- and E7-dependent p53 and pRb degradations. All these data suggest that OI could be explored for its chemotherapeutic potential in cervical cancer cells with HPV-integrated genomes.
  2. Fulltext Direct Resazurin Microplate Assay in Drug Susceptibility Testing of Smear-Positive Sputum Samples against Mycobacterium tuberculosis
    Jamil NIN, Wahab WNAWA, Ali IA, Yahaya ML
    Malays J Med Sci, 2018 Nov;25(6):59-66.
    PMID: 30914879 DOI: 10.21315/mjms2018.25.6.6
    BACKGROUND: A new direct microplate-based colorimetric drug susceptibility test that omits the initial isolation of Mycobacterium tuberculosis from sputum specimens was evaluated.

    METHODS: A total of 51 M. tuberculosis acid fast bacilli (AFB) smear-positive sputum specimens were inoculated directly into drug-free and serial dilutions of drug-containing Middlebrook 7H9 broth media. With this direct resazurin micro plate assay (REMA) method, resazurin dye was used as a growth indicator in microplate wells. The minimum inhibitory concentrations (MIC) of isoniazid (INH) and rifampicin (RIF) were compared with those of the 'gold standard' absolute concentration method (ACM). The turnaround time (TAT) of the direct REMA and the ACM were also determined.

    RESULTS: At the selected cut-off points (INH: 0.0625 μg/mL; RIF: 0.125 μg/mL), good drug susceptibility test results were obtained for INH and RIF with an average sensitivity, specificity and accuracy of 90%, 100% and 97%, respectively, with a TAT of 15 days. The REMA method also correctly classified the resistant isolates with positive predictive values of 95% and negative predictive values of 98% for the two drugs.

    CONCLUSIONS: The direct REMA was reliable in routine diagnostic laboratories for the drug susceptibility testing of M. tuberculosis and the rapid detection of multi-drug-resistant tuberculosis.

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